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| affinity chromatographySummarySummary: A chromatographic technique that utilizes the ability of biological molecules to bind to certain ligands specifically and reversibly. It is used in protein biochemistry. (McGraw-Hill Dictionary of Scientific and Technical Terms, 4th ed) Top Publications
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Structural analysis and classification of native proteins from E. coli commonly co-purified by immobilised metal affinity chromatography
Victor Martin Bolanos-Garcia
Department of Biochemistry, University of Cambridge, 80 Tennis Court Road, Cambridge CB2 1GA, England
Biochim Biophys Acta 1760:1304-13. 2006Immobilised metal affinity chromatography (IMAC) is the most widely used technique for single-step purification of recombinant proteins...Identification of putative serum glycoprotein biomarkers for human lung adenocarcinoma by multilectin affinity chromatography and LC-MS/MS
Sun Hee Heo
Department of Biochemistry, School of Dentistry, Kyungpook National University, Daegu, Korea
Proteomics 7:4292-302. 2007..from the sera of three healthy individuals and three lung adenocarcinoma patients using multilectin affinity chromatography. The recovered glycoproteins were subjected to treatment with peptide-N-glycosidase F (PNGase F) and in-..Functional organization of the yeast proteome by systematic analysis of protein complexes
Anne Claude Gavin
Cellzome AG, Meyerhofstrasse 1, 69117 Heidelberg, Germany
Nature 415:141-7. 2002..This higher-order map contains fundamental biological information and offers the context for a more reasoned and informed approach to drug discovery...An efficient tandem affinity purification procedure for interaction proteomics in mammalian cells
Tilmann Burckstummer
Research Center for Molecular Medicine CeMM, Lazarettgasse 19 3, 1090 Vienna, Austria
Nat Methods 3:1013-9. 2006..Using the well-characterized Ku70-Ku80 protein complex as an example, we identified both core elements as well as new candidate effectors...Enrichment and analysis of nonenzymatically glycated peptides: boronate affinity chromatography coupled with electron-transfer dissociation mass spectrometry
Qibin Zhang
Biological Sciences Division and Environmental Molecular Sciences Laboratory, Pacific Northwest National Laboratory, Richland, Washington 99352, USA
J Proteome Res 6:2323-30. 2007..In this report, phenylboronate affinity chromatography was used in a two-step enrichment scheme to selectively isolate first glycated proteins and then glycated,..Oligosaccharide specificity of galectins: a search by frontal affinity chromatography
Jun Hirabayashi
Department of Biological Chemistry, Faculty of Pharmaceutical Sciences, Teikyo University, Sagamiko, Kanagawa 199 0195, Japan
Biochim Biophys Acta 1572:232-54. 2002..For this purpose, we recently reinforced a conventional system for frontal affinity chromatography (FAC) [J. Chromatogr., B, Biomed. Sci. Appl. 771 (2002) 67-87]...Specific phosphopeptide enrichment with immobilized titanium ion affinity chromatography adsorbent for phosphoproteome analysis
Houjiang Zhou
National Chromatographic R and A Center, Dalian Institute of Chemical Physics, Chinese Academy of Sciences, Dalian 116023, China
J Proteome Res 7:3957-67. 2008..Here, we presented a new method termed as immobilized titanium ion affinity chromatography (Ti (4+)-IMAC) for enriching phosphopeptides...Highly efficient tandem affinity purification of trypanosome protein complexes based on a novel epitope combination
Bernd Schimanski
Department of Genetic and Developmental Biology, University of Connecticut Health Center, 263 Farmington Ave, Farmington, CT 06030 3301, USA
Eukaryot Cell 4:1942-50. 2005..Specifically, the calmodulin affinity chromatography step proved to be inefficient...Characterization of cDNAs encoding the p44 and p35 subunits of human translation initiation factor eIF3
K L Block
Department of Biological Chemistry, School of Medicine, University of California, Davis, California 95616, USA
J Biol Chem 273:31901-8. 1998..Possession of cloned cDNAs encoding all 10 subunits of eIF3 provides the tools necessary to elucidate the functions of the individual subunits and the structure of the eIF3 complex...Molecular identification of a novel candidate sorting receptor purified from human brain by receptor-associated protein affinity chromatography
C M Petersen
Department of Medical Biochemistry, University of Aarhus, 8000 Aarhus C, Denmark
J Biol Chem 272:3599-605. 1997..95-kDa membrane glycoprotein, designated gp95/sortilin, was purified from human brain extracts by RAP affinity chromatography and cloned in a human cDNA library...An alternative tandem affinity purification strategy applied to Arabidopsis protein complex isolation
Vicente Rubio
Department of Molecular, Cellular, and Developmental Biology, Yale University, New Haven, CT 06520 8104, USA
Plant J 41:767-78. 2005..Taken together, the data demonstrate the suitability of the TAPa system to allow efficient multi-protein complex isolation from stably transformed Arabidopsis...Isolation and identification of the third subunit of mammalian DNA polymerase delta by PCNA-affinity chromatography of mouse FM3A cell extracts
P Hughes
Centre National de la Recherche Scientifique CNRS, UPR9044, Institut de Recherches sur le Cancer, 7 rue Guy Moquet BP 8, 94801 Villejuif, France
Nucleic Acids Res 27:2108-14. 1999..We discuss the implications of our findings within the current framework of our understanding of DNA replication...Synergistic activation of transcription by CBP and p53
W Gu
Laboratory of Biochemistry and Molecular Biology, The Rockefeller University, New York 10021, USA
Nature 387:819-23. 1997....Immobilized metal affinity chromatography revisited: pH/acid control toward high selectivity in phosphoproteomics
Chia Feng Tsai
Department of Chemistry, National Taiwan Normal University, Taipei, Taiwan
J Proteome Res 7:4058-69. 2008..Furthermore, the simple and reproducible IMAC protocol can be adapted to other solid supports, fully automated or manual, for large-scale identification of the vastly under-explored phosphoproteome...A physical and functional map of the human TNF-alpha/NF-kappa B signal transduction pathway
Tewis Bouwmeester
Cellzome AG, Meyerhofstrasse 1, 69117 Heidelberg, Germany
Nat Cell Biol 6:97-105. 2004..This systems approach provides significant insight into the logic of the TNF-alpha/NF-kappa B pathway and is generally applicable to other pathways relevant to human disease...Quantitative interaction proteomics using mass spectrometry
Alexander Wepf
Institute of Molecular Systems Biology, Swiss Federal Institute of Technology, Eidgenossische Technische Hochschule Zurich, Wolfgang Pauli Strasse 16, Zurich, Switzerland
Nat Methods 6:203-5. 2009..We used this method to quantitatively analyze interaction stoichiometries in the human protein phosphatase 2A network...Purification of Shiga toxin and Shiga-like toxins I and II by receptor analog affinity chromatography with immobilized P1 glycoprotein and production of cross-reactive monoclonal antibodies
A Donohue Rolfe
Department of Medicine, New England Medical Center, Boston, Massachusetts
Infect Immun 57:3888-93. 1989Shiga toxin from Shigella dysenteriae 60R was purified to homogeneity by a novel one-step receptor analog affinity chromatography method...Mapping multiprotein complexes by affinity purification and mass spectrometry
Mark O Collins
Proteomic Mass Spectrometry, The Wellcome Trust Sanger Institute, Hinxton, Cambridge CB10 1SA, UK
Curr Opin Biotechnol 19:324-30. 2008..Concomitant developments in MS, including high accuracy and high-throughput instrumentation together with quantitative MS methods, have facilitated large-scale and comprehensive analysis of multiprotein complexes...Identification of 40LoVe, a Xenopus hnRNP D family protein involved in localizing a TGF-beta-related mRNA during oogenesis
Kevin Czaplinski
EMBL Gene Expression Programme, Meyerhofstrasse 1, Heidelberg 69117, Germany
Dev Cell 8:505-15. 2005..Using an RNA affinity chromatography approach, we investigated localization RNP formation on the vegetal localization element (VLE) of the ..Protein-protein interaction assays: eliminating false positive interactions
Tuan N Nguyen
Department of Chemistry and Biochemistry, University of Colorado, Boulder, Colorado 80309, USA
Nat Methods 3:135-9. 2006Purification of Drosophila DNA polymerase zeta by REV1 protein-affinity chromatography
Ryo Takeuchi
Department of Applied Biological Science, Faculty of Science and Technology, Tokyo University of Science, Noda shi, Chiba ken 278 8510, Japan
Biochem J 382:535-43. 2004..The present study is the first report about characterization of purified pol zeta from multicellular organisms, and the second concerning the characterization of yeast pol zeta...Composition of the synaptic PSD-95 complex
Ayse Dosemeci
Laboratory of Neurobiology, NINDS, National Institutes of Health, Bethesda, Maryland 20892, USA
Mol Cell Proteomics 6:1749-60. 2007..Another co-purifying protein (Q8BZM2) with two sterile alpha motif domains may represent a novel structural core element of the PSD...Purification of prostate-specific membrane antigen using conformational epitope-specific antibody-affinity chromatography
Tiancheng Liu
Department of Chemistry and Biochemistry, San Francisco State University, 1600 Holloway Ave, San Francisco, CA 94132, USA
Protein Expr Purif 49:251-5. 2006..optimized the purification of native PSMA from LNCaP cells using conformational epitope-specific antibody-affinity chromatography. Western blot analysis and an HPLC-based enzymatic activity assay were used to compare the yield and ..Affinity chromatography with collapsibly tethered ligands
Hidenori Yamanaka
Department of Research and Development, Amersham Biosciences K K, 3 25 1 Hyakunincho, Shinjuku ku, Tokyo 169 0073, Japan
Anal Chem 75:1658-63. 2003We introduce a novel affinity chromatography mode in which affinity ligands are secured to the media surface via collapsible tethers...Preparation of monodisperse immobilized Ti(4+) affinity chromatography microspheres for specific enrichment of phosphopeptides
Zhiyuan Yu
National Chromatographic R and A Center, Dalian Institute of Chemical Physics, Chinese Academy of Sciences, Dalian 116023, China
Anal Chim Acta 636:34-41. 2009This study presented an approach to prepare monodisperse immobilized Ti(4+) affinity chromatography (Ti(4+)-IMAC) microspheres for specific enrichment of phosphopeptides in phosphoproteome analysis...Current strategies for the use of affinity tags and tag removal for the purification of recombinant proteins
Jose Arnau
Unizyme Laboratories A S, Dr Neergaards vej 17, DK 2970 Hørsholm, Denmark
Protein Expr Purif 48:1-13. 2006..A comparison of the yield of standard vs. affinity purification together with an example of tag removal using TAGZyme is also included...Synthesis of conjugates of 6-aminophenanthridine and guanabenz, two structurally unrelated prion inhibitors, for the determination of their cellular targets by affinity chromatography
Fabienne Gug
Laboratoire de Chimie Organique 2, INSERM U648, Universite Paris Descartes, 4 avenue de l Observatoire, 75270 Paris Cedex 06, France
Bioconjug Chem 21:279-88. 2010..Here, we show, using competition experiments with Quinacrine (QC) and Chlorpromazine (CPZ), two other antiprion drugs, that QC, but not CPZ, may also directly target the rRNA...Identification of glutathione sulfotransferase-pi (GSTP1) as a new resveratrol targeting protein (RTP) and studies of resveratrol-responsive protein changes by resveratrol affinity chromatography
Tze Chen Hsieh
Department of Biochemistry and Molecular Biology, New York Medical College, Valhalla, NY 10595, USA
Anticancer Res 28:29-36. 2008..The bioactivities of resveratrol may be attributed to qualitative and quantitative differences in its cell-type-specific interaction and binding with its cellular targets, denoted as resveratrol targeting proteins (RTPs)...Frontal affinity chromatography analysis of constructs of DC-SIGN, DC-SIGNR and LSECtin extend evidence for affinity to agalactosylated N-glycans
Rikio Yabe
Research Center for Medical Glycoscience, National Institute of Advanced Industrial Science and Technology AIST, Tsukuba, Ibaraki, Japan
FEBS J 277:4010-26. 2010..DC-SIGNR) and liver and lymph node sinusoidal endothelial cell C-type lectin (LSECtin) using frontal affinity chromatography (FAC)...Enrichment and characterization of phosphopeptides by immobilized metal affinity chromatography (IMAC) and mass spectrometry
Tine E Thingholm
Department of Biochemistry and Molecular Biology, University of Southern Denmark, Odense, Denmark
Methods Mol Biol 527:47-56, xi. 2009The combination of immobilized metal affinity chromatography (IMAC) and mass spectrometry is a widely used technique for enrichment and sequencing of phosphopeptides...Online immobilized metal affinity chromatography/mass spectrometric analysis of changes elicited by cCMP in the murine brain phosphoproteome
S Ding
Department of Environmental and Molecular Biosciences, Swansea University, SOTEAS, Singleton Park, Swansea SA2 8PP, UK
Rapid Commun Mass Spectrom 22:4129-38. 2008An automated online immobilized metal affinity chromatography/high-performance liquid chromatography mass spectrometric (IMAC-HPLC/MS/MS) method was developed to study cytidine 3',5'-cyclic monophosphate (cCMP)-specific protein ..Displacement affinity chromatography of protein phosphatase one (PP1) complexes
Greg B G Moorhead
Department of Biological Sciences, University of Calgary, 2500 University Dr, NW Calgary, AB T2N 1N4, Canada
BMC Biochem 9:28. 2008..PP1 is localized to its site of action by interacting with targeting or regulatory proteins, a majority of which contains a primary docking site referred to as the RVXF/W motif...Analysis of desiccation-induced candidate phosphoproteins from Craterostigma plantagineum isolated with a modified metal oxide affinity chromatography procedure
Horst Rohrig
Institute of Molecular Physiology and Biotechnology of Plants, University of Bonn, Bonn, Germany
Proteomics 8:3548-60. 2008..Possible functions of selected proteins are discussed in the context of the de-/rehydration cycle...Enrichment and analysis of phosphopeptides under different experimental conditions using titanium dioxide affinity chromatography and mass spectrometry
Uma K Aryal
National Research Council, Plant Biotechnology Institute, 110 Gymnasium Place, Saskatoon, Saskatchewan, Canada S7N 0W9
Rapid Commun Mass Spectrom 24:219-31. 2010Titanium dioxide metal oxide affinity chromatography (TiO(2)-MOAC) is widely regarded as being more selective than immobilized metal-ion affinity chromatography (IMAC) for phosphopeptide enrichment...Purification of human haptoglobin 1-1, 2-1, and 2-2 using monoclonal antibody affinity chromatography
Chi Feng Tseng
Department of Biological Science and Technology, Research Institute of Biochemical Engineering, National Chiao Tung University, Hsinchu, Taiwan, ROC
Protein Expr Purif 33:265-73. 2004..Circular dichroic spectra showed that the alpha-helical content of Hp 1-1 (29%) was higher than that of Hp 2-1 (22%), and 2-2 (21%). The structural difference with respect to its clinical relevance is discussed...85-kDa protein of Trypanosoma cruzi purified by affinity chromatography used in the multiple antigen binding assay (MABA) for the diagnosis of T. cruzi infection in a Venezuelan rural community
Mariolga Berrizbeitia
Postgrado en Biología Aplicada, Nucleo de Sucre, Universidad de Oriente, Cumana, Venezuela
Parasitol Res 106:1127-34. 2010..cruzi infection in this region of Venezuela...Screening of protein-ligand interactions using dynamic protein-affinity chromatography solid-phase extraction-liquid chromatography-mass spectrometry
Niels Jonker
VU University Amsterdam, Faculty of Sciences, Department of Chemistry, Biomolecular Analysis Group, De Boelelaan 1083, 1081 HV Amsterdam, The Netherlands
J Chromatogr A 1205:71-7. 2008..The system presented, dynamic protein-affinity chromatography solid-phase extraction (DPAC-SPE), overcomes the limitations of the existing methods by performing an ..Immunopurification of polyribosomal complexes of Arabidopsis for global analysis of gene expression
Maria Eugenia Zanetti
Department of Botany and Plant Science, Center for Plant Cell Biology, University of California, Riverside, California 92521 0124, USA
Plant Physiol 138:624-35. 2005..This technology can be extended to evaluation of mRNA populations at the cell- or tissue-specific level by regulation of the tagged RP with distinct promoters...Affinity chromatography: general methods
Marjeta Urh
Promega Corporation, Madison, Wisconsin, USA
Methods Enzymol 463:417-38. 2009b>Affinity chromatography is one of the most diverse and powerful chromatographic methods for purification of a specific molecule or a group of molecules from complex mixtures...Monoacetylcurcumin: a new inhibitor of eukaryotic DNA polymerase lambda and a new ligand for inhibitor-affinity chromatography
Yoshiyuki Mizushina
Laboratory of Food and Nutritional Sciences, Department of Nutritional Science, Kobe Gakuin University, Nishi ku, Kobe, Hyogo 651 2180, Japan
Biochem Biophys Res Commun 337:1288-95. 2005..We then made a monoacetylcurcumin-conjugated column with epoxy-activated Sepharose 6B. In the column, pol lambda of full length was selectively adsorbed and eluted...Analysis of glycopeptides using lectin affinity chromatography with MALDI-TOF mass spectrometry
Kazutosi Kubota
Research Team for Functional Genomics and Research Team for Molecular Biomarkers, Tokyo Metropolitan Institute of Gerontology, Foundation for Research on Aging and Promotion of Human Welfare, 35 2 Sakae cho, Itabashi ku, Tokyo 173 0015, Japan
Anal Chem 80:3693-8. 2008..Taken together, the combination of lectin column chromatography, MALDI-TOF MS, and MALDI-QIT-TOF MS ( n ) provides an easy way for the structural estimation of glycans and the rapid analysis of glycoproteomics...Cathepsin B-like cysteine proteases confer intestinal cysteine protease activity in Haemonchus contortus
S Shompole
Department of Veterinary Microbiology and Pathology, Washington State University, Pullman 99164 7040, USA
J Biol Chem 276:2928-34. 2001..Data indicate that CBLs are responsible for cysteine protease activity described from H. contortus intestine. Results also support a role of CBLs in nutrient digestion...Expression, biotinylation and purification of a biotin-domain peptide from the biotin carboxy carrier protein of Escherichia coli acetyl-CoA carboxylase
A Chapman-Smith
Department of Biochemistry, University of Adelaide, Australia
Biochem J 302:881-7. 1994..The identify of the purified biotin-domain peptide was confirmed by N-terminal sequence analysis, amino acid analysis and m.s. The domain was readily produced and purified in sufficient quantities for n.m.r. structural analysis...Characterization of a mammalian homolog of the Escherichia coli MutY mismatch repair protein
J P McGoldrick
Department of Biological Chemistry, School of Medicine, University of Maryland at Baltimore, 21201
Mol Cell Biol 15:989-96. 1995..The eukaryotic MYH may be involved in the major repair of both replication errors and oxidative damage to DNA, the same functions as those of the E. coli MutY protein...The mitotic apparatus-associated 51-kDa protein from sea urchin eggs is a GTP-binding protein and is immunologically related to yeast polypeptide elongation factor 1 alpha
K Ohta
Department of Biophysics and Biochemistry, Faculty of Science, University of Tokyo, Japan
J Biol Chem 265:3240-7. 1990..The results indicate that the mitotic apparatus-associated 51-kDa protein is a GTP-binding protein and suggest that it is structurally and functionally related to yeast EF-1 alpha...Retention and separation of adenosine and analogues by affinity chromatography with an aptamer stationary phase
Q Deng
Department of Chemistry, University of Florida, Gainesville 32611, USA
Anal Chem 73:5415-21. 2001..The aptamer medium was packed into fused-silica capillaries (50-150-microm i.d.) to form affinity chromatography columns...Single-step Strep-tag purification for the isolation and identification of protein complexes from mammalian cells
Melissa R Junttila
Centre for Biotechnology, University of Turku and Abo Akademi University, Turku, Finland
Proteomics 5:1199-203. 2005..By using the Strep-tag affinity chromatography method, enabling fast and simple one-step purification, coupled with competitive elution under ..O-linked N-acetylglucosamine proteomics of postsynaptic density preparations using lectin weak affinity chromatography and mass spectrometry
Keith Vosseller
Mass Spectrometry Facility, Department of Pharmaceutical Chemistry, University of California, San Francisco, California 94143, USA
Mol Cell Proteomics 5:923-34. 2006..for direct enrichment and identification of in vivo O-GlcNAc-modified peptides through lectin weak affinity chromatography (LWAC) and mass spectrometry...Robust phosphoproteomic profiling of tyrosine phosphorylation sites from human T cells using immobilized metal affinity chromatography and tandem mass spectrometry
Laurence M Brill
Genomics Institute of the Novartis Research Foundation, 10675 John Jay Hopkins Drive, San Diego, California 92121, USA
Anal Chem 76:2763-72. 2004..2003, 100, 443-448). The technology platform included the use of immunoprecipitation, immobilized metal affinity chromatography (IMAC), liquid chromatography, and tandem mass spectrometry...Cell-free translation reconstituted with purified components
Y Shimizu
Department of Chemistry and Biotechnology, Graduate School of Engineering, The University of Tokyo, Bldg FSB401, Kasiwanoha, Kashiwa, Chiba Prefecture 277-8562, Japan
Nat Biotechnol 19:751-5. 2001..The protein products were easily purified within 1 h using affinity chromatography to remove the tagged protein factors...Combination of abundant protein depletion and multi-lectin affinity chromatography (M-LAC) for plasma protein biomarker discovery
Tatiana Plavina
Biogen Idec Inc, Boston, MA 02142, USA
J Proteome Res 6:662-71. 2007..The method consists of depletion of albumin and IgG and multi-lectin affinity chromatography (M-LAC), followed by nanoLC-MS/MS analysis of digested proteins and label-free comparative quantitation ..Systematic comparison of oligosaccharide specificity of Ricinus communis agglutinin I and Erythrina lectins: a search by frontal affinity chromatography
Yoko Itakura
Lectin Application and Analysis Team, Research Center for Medical Glycoscience, National Institute of Advanced Industrial Science and Technology, Tsukuba Central 2, 1 1 1, Umezono, Tsukuba, Ibaraki 305 8568
J Biochem 142:459-69. 2007..For this purpose, we examined by means of frontal affinity chromatography over 100 lectins which have similar sugar-binding specificities to that of RCA120...Multi-component immunoaffinity subtraction chromatography: an innovative step towards a comprehensive survey of the human plasma proteome
Rembert Pieper
Large Scale Biology Corporation, Germantown, Maryland, USA
Proteomics 3:422-32. 2003..This facilitates automated chromatographic processing of plasma samples in high throughput, which is desirable in proteomic disease marker discovery projects...Analysis of drug-protein binding by ultrafast affinity chromatography using immobilized human serum albumin
Rangan Mallik
Chemistry Department, University of Nebraska, Lincoln, NE 68588 0304, USA
J Chromatogr A 1217:2796-803. 2010..These properties make this technique appealing for use in drug-binding studies and in the high-throughput screening of new drug candidates...Approaches to the study of N-linked glycoproteins in human plasma using lectin affinity chromatography and nano-HPLC coupled to electrospray linear ion trap--Fourier transform mass spectrometry
Yonghui Wang
Barnett Institute, Northeastern University, Boston, MA 02115, USA
Glycobiology 16:514-23. 2006In this publication, we will describe the combination of lectin affinity chromatography with nano high performance liquid chromatography (HPLC) coupled to a linear ion trap Fourier transform mass spectrometer (capillary LC-LTQ/FTMS) to ..RNA affinity for molecular L-histidine; genetic code origins
Irene Majerfeld
Molecular, Cellular and Developmental Biology, University of Colorado, Boulder, CO 80309 0347, USA
J Mol Evol 61:226-35. 2005..This site is probably the simplest structure that can meet our histidine affinity selection, which strengthens experimental support for a "stereochemical" origin of the genetic code...Lectin affinity chromatography as a tool to differentiate endogenous and recombinant erythropoietins
L Franco Fraguas
Catedra de Bioquimica, Departamento de Biociencias, Facultad de Quimica, CC 1157, Montevideo, Uruguay
J Chromatogr A 1212:82-8. 2008This work exploits the combination of the lectin affinity chromatography (LAC) with an ultra-sensitive immunochromatographic assay to differentiate several types of erythropoietin (EPO)...High-throughput protein purification using an automated set-up for high-yield affinity chromatography
Johanna Steen
School of Biotechnology, Division of Molecular Biotechnology, AlbaNova University Center, KTH, Royal Institute of Technology, Roslagsvägen 30 B, 106 91 Stockholm, Sweden
Protein Expr Purif 46:173-8. 2006..5h. The method is based on affinity chromatography and has been set up on an instrument that utilizes positive pressure for liquid transfer through columns...A novel tandem affinity purification strategy for the efficient isolation and characterisation of native protein complexes
Christian Johannes Gloeckner
Institute of Human Genetics, GSF National Research Center for Environment and Health, Neuherberg, Germany
Proteomics 7:4228-34. 2007....Glutamine synthetase, hemoglobin alpha-chain, and macrophage migration inhibitory factor binding to amyloid beta-protein: their identification in rat brain by a novel affinity chromatography and in Alzheimer's disease brain by immunoprecipitation
R Oyama
Department of Neuropathology, Faculty of Medicine, University of Tokyo, Japan
Biochim Biophys Acta 1479:91-102. 2000..Our results suggest that these binding proteins also associate with Abeta, leading to the clearance or the accumulation of Abeta and the neuronal cell damage in human brain...SATB1 targets chromatin remodelling to regulate genes over long distances
Dag Yasui
Life Sciences Division, Lawrence Berkeley National Laboratory, University of California, Berkeley 94720, USA
Nature 419:641-5. 2002..SATB1 defines a class of transcriptional regulators that function as a 'landing platform' for several chromatin remodelling enzymes and hence regulate large chromatin domains...Rapid enrichment and analysis of yeast phosphoproteins using affinity chromatography, 2D-PAGE and peptide mass fingerprinting
Vasso Makrantoni
Centre for Biomolecular Science, University of St Andrews, The North Haugh, St Andrews, Fife KY16 9ST, UK
Yeast 22:401-14. 2005..Further validating the method, growth of yeast in the presence of sorbic acid resulted in altered phosphorylation of 17 proteins, 13 of which had prior published evidence that they were phosphoproteins or had ATP binding activity...Identification of intracellular targets of small molecular weight chemical compounds using affinity chromatography
Damien Guiffant
CNRS, Molecules and Therapeutic Targets Laboratory, Station Biologique, Roscoff, France
Biotechnol J 2:68-75. 2007..Here we discuss the use of immobilized drugs as affinity chromatography matrices to purify and identify their bona fide intracellular targets...High-throughput analysis of lectin-oligosaccharide interactions by automated frontal affinity chromatography
Sachiko Nakamura-Tsuruta
AIST, Research Center for Glycoscience, Ibaraki, Japan
Methods Enzymol 415:311-25. 2006Frontal affinity chromatography (FAC) is a quantitative method that enables sensitive and reproducible measurements of interactions between lectins and oligosaccharides...Tandem affinity purification of functional TAP-tagged proteins from human cells
Juraj Gregan
Department of Chromosome Biology, Max F Perutz Laboratories, University of Vienna, Dr Bohr Gasse 1, 1030 Vienna, Austria
Nat Protoc 2:1145-51. 2007..The purification protocol can be completed in 1 day and it should be applicable to other proteins...Adsorptive refolding of histidine-tagged glutathione S-transferase using metal affinity chromatography
Matthew H Hutchinson
Department of Chemical Engineering, University of Cambridge, Pembroke Street, Cambridge CB2 3RA, UK
J Chromatogr A 1128:125-32. 2006..His6-tagged glutathione S-transferase (GST-(His6)) was refolded while it was adsorbed in a metal affinity chromatography column...Immobilized metal affinity chromatography without chelating ligands: purification of soybean trypsin inhibitor on zinc alginate beads
Munishwar N Gupta
Chemistry Department, Indian Institute of Technology, Delhi, Hauz Khas, New Delhi 110016, India
Biotechnol Prog 18:78-81. 2002Immobilized metal affinity chromatography (IMAC) is a widely used technique for bioseparation of proteins in general and recombinant proteins with polyhistidine fusion tags in particular...Enrichment of multiphosphorylated peptides by immobilized metal affinity chromatography using Ga(III)- and Fe(III)-complexes
Corinna Sykora
Institute of Bioanalytical Chemistry, Centre for Biotechnology and Biomedicine BBZ, Faculty of Chemistry and Mineralogy, Leipzig University, Deutscher Platz 5, 04103 Leipzig, Germany
Protein Pept Lett 14:489-96. 2007..A common approach to analyse these modifications is to enrich phosphopeptides by immobilized metal affinity chromatography (IMAC) prior to mass spectrometric analysis...Site-directed biotinylation of antibodies for controlled immobilization on solid surfaces
Il Hoon Cho
Program for Bio Microsystem Technology, Korea University, 1, 5 Ka, Anam Dong, Sungbuk Ku, Seoul 136 701, Korea
Anal Biochem 365:14-23. 2007..Preparations of both the intact antibody and the F(ab')(2) showed consistently enhanced detection capabilities that were 2.6 and 20 times that of the control, respectively...Histidines in affinity tags and surface clusters for immobilized metal-ion affinity chromatography of trimeric tumor necrosis factor alpha
V Gaberc-Porekar
National Institute of Chemistry, Hajdrihova, Ljubljana, Slovenia
J Chromatogr A 852:117-28. 1999In order to achieve efficient IMAC (immobilized metal-ion affinity chromatography) purification of tumor necrosis factor alpha (TNF-alpha) and its analogs by a common chromatographic procedure, we tested four histidine-rich affinity tags ..Affinity chromatography: a review of clinical applications
D S Hage
Department of Chemistry, 738 Hamilton Hall, University of Nebraska, Lincoln, NE 68588 0304 Fax 402 472 9402, USA
Clin Chem 45:593-615. 1999b>Affinity chromatography is a type of liquid chromatography that makes use of biological-like interactions for the separation and specific analysis of sample components...An improved strategy for high-level production of TEV protease in Escherichia coli and its purification and characterization
Lei Fang
The State Key Laboratory of Pharmaceutical Biotechnology and Department of Biochemistry, College of Life Science, Nanjing University, Nanjing 210093, PR China
Protein Expr Purif 51:102-9. 2007..When further purified by Ni(2+) affinity chromatography, 65mg of His6-TEV was isolated with purity over 95% from 1L of culture...Protein composition and electron microscopy structure of affinity-purified human spliceosomal B complexes isolated under physiological conditions
Jochen Deckert
Department of Cellular Biochemistry, Max Planck Institute for Biophysical Chemistry, Am Fassberg 11, 37077 Gottingen, Germany
Mol Cell Biol 26:5528-43. 2006..Furthermore, the spliceosomal complexes isolated here are well suited for complementation studies with purified proteins to dissect factor requirements for spliceosome activation and splicing catalysis...Engineering green fluorescent protein as a dual functional tag
Rosanto I Paramban
Department of Molecular Biosciences and Bioengineering, University of Hawaii at Manoa, Honolulu, Hawaii 96822, USA
Biotechnol Bioeng 86:687-97. 2004..The variants were produced in Escherichia coli and purified using immobilized metal affinity chromatography (IMAC). The purification efficiencies by IMAC for all variants were found to be comparable...Enrichment of phosphorylated proteins and peptides from complex mixtures using metal oxide/hydroxide affinity chromatography (MOAC)
Florian Wolschin
Max Planck Institute of Molecular Plant Physiology, Potsdam, Germany
Proteomics 5:4389-97. 2005A novel method termed metal oxide affinity chromatography (MOAC) of enriching for phosphorylated proteins and peptides based on the affinity of the phosphate group for Al(OH)(3) is presented here...High-throughput purification and quality assurance of Arabidopsis thaliana proteins for eukaryotic structural genomics
Won Bae Jeon
The Center for Eukaryotic Structural Genomics, University of Wisconsin Madison, 433 Bobcock Drive, 53706 1549, USA
J Struct Funct Genomics 6:143-7. 2005..Using an automated AKTApurifier system, fusion proteins were initially purified by immobilized metal affinity chromatography (IMAC)...Tropoelastin interacts with cell-surface glycosaminoglycans via its COOH-terminal domain
Thomas J Broekelmann
Department of Cell Biology and Physiology, Washington University School of Medicine, St Louis, Missouri 63110, USA
J Biol Chem 280:40939-47. 2005..Heparin affinity chromatography combined with cell adhesion assays identified the last 17 amino acids as the sequence element at the ..A high-capacity RNA affinity column for the purification of human IRP1 and IRP2 overexpressed in Pichia pastoris
Charles R Allerson
Cell Biology and Metabolism Branch, National Institute of Child Health and Human Development, National Institutes of Health, Bethesda, Maryland 20892, USA
RNA 9:364-74. 2003..liter of culture) of each human IRP in the yeast Pichia pastoris and large-scale purification using RNA affinity chromatography. Despite the utility of RNA affinity chromatography in the isolation of RNA-binding proteins, current ..Membrane-associated, boron-interacting proteins isolated by boronate affinity chromatography
Monika A Wimmer
Institute of Crop Science and Resource Conservation, University of Bonn, Bonn, Germany
Plant Cell Physiol 50:1292-304. 2009..microsomal preparations of arabidopsis (Arabidopsis thaliana) and maize (Zea mays) using phenylboronate affinity chromatography, subsequently separated by two-dimensional gel electrophoresis and identified using MALDI-TOF (matrix-..Glycoproteomic analysis of embryonic stem cells: identification of potential glycobiomarkers using lectin affinity chromatography of glycopeptides
Gerardo Alvarez-Manilla
Complex Carbohydrate Research Center, University of Georgia, Athens, Georgia 30602, USA
J Proteome Res 9:2062-75. 2010..are part of a very complex proteome, has involved the use of fractionation techniques such as lectin affinity chromatography. In this study, the glycoproteomic characterization of pluripotent murine embryonic stem cells (ES) and ..Protection studies in sheep using affinity-purified and recombinant cysteine proteinases of adult Haemonchus contortus
Diane L Redmond
Moredun Research Institute, International Research Centre, Pentlands Science Park, Penicuik, Midlothian EH260PZ, Scotland, UK
Vaccine 22:4252-61. 2004..In this study, the cysteine proteinases of adult H. contortus TSBP were specifically purified by affinity chromatography using recombinant H. contortus cystatin, a potent cysteine proteinase inhibitor. Although only 1-1...Purification and characterization of three toxins and two agglutinins from Abrus precatorius seed by using lactamyl-Sepharose affinity chromatography
R Hegde
Department of Biochemistry, Indian Institute of Science, Bangalore
Anal Biochem 194:101-9. 1991..two agglutinins, APA-I and -II, were purified from the seeds of Abrus precatorius by lactamyl-Sepharose affinity chromatography followed by gel filtration and DEAE-Sephacel column chromatography...An integrated workflow for charting the human interaction proteome: insights into the PP2A system
Timo Glatter
Institute of Molecular Systems Biology, ETH Zurich, Zurich, Switzerland
Mol Syst Biol 5:237. 2009..These results show that the presented analytical process will substantially advance throughput and reproducibility in future systematic AP-MS studies on human protein complexes...Purification of antibodies by affinity chromatography
Klaus Huse
Institute of Molecular Biotechnology, Jena, Germany
J Biochem Biophys Methods 51:217-31. 2002..The purification strategies for antibodies by interaction with affinity ligands such as antibodies and Fe receptors or low molecular weight compounds are described...A novel method to quantify sphingosine 1-phosphate by immobilized metal affinity chromatography (IMAC)
Yong Moon Lee
Center for Vascular Biology, Department of Cell Biology, University of Connecticut Health Center, Farmington, CT 06030 3505, USA
Prostaglandins Other Lipid Mediat 84:154-62. 2007..to selectively enrich S1P and dihydro-S1P from biological samples by the Fe(3+) gel immobilized metal affinity chromatography (IMAC)...Frontal affinity chromatography: sugar-protein interactions
Hiroaki Tateno
Research Center for Medical Glycoscience, National Institute of Advanced Industrial Science and Technology, AIST Tsukuba Central 2, 1 1 1 Umezono, Ibaraki 305 8568, Japan
Nat Protoc 2:2529-37. 2007Frontal affinity chromatography using fluorescence detection (FAC-FD) is a versatile technique for the precise determination of dissociation constants (Kd) between glycan-binding proteins (lectins) and fluorescent-labeled glycans...Isolation of new pregnancy-associated glycoproteins from water buffalo (Bubalus bubalis) placenta by Vicia villosa affinity chromatography
O Barbato
Department of Biopathological Veterinary Science, Faculty of Veterinary Medicine, University of Perugia 06126, Italy
Res Vet Sci 85:457-66. 2008..the water buffalo PAG (wbPAG) from these solutions are enriched by Vicia villosa agarose (VVA) affinity chromatography. As determined by western blotting with anti-PAG sera, the apparent molecular masses of the ..Identification of dengue virus binding proteins using affinity chromatography
Supranee Upanan
Molecular Pathology Laboratory, Institute of Molecular Biology and Genetics, Mahidol University, Salaya Campus, 25 25 Phuttamonthol Sai 4, Salaya, Nakorn Pathom 73170, Thailand
J Virol Methods 151:325-8. 2008..This result is consistent with earlier work and shows that while affinity chromatography may provide a useful adjunct to VOPBA based studies particularly in cases where proteins maybe sensitive ..Analysis of amadori peptides enriched by boronic acid affinity chromatography
Andrej Frolov
Institute of Bioanalytical Chemistry, Center for Biotechnology and Biomedicine, Faculty of Chemistry and Mineralogy, Leipzig University, Leipzig, Germany
Ann N Y Acad Sci 1126:253-6. 2008..Here, we describe a universal enrichment procedure for glycated peptides using boronic acid affinity chromatography in the first dimension followed by reversed-phase chromatography, coupled either online to electrospray ..Glycoproteomics of plasma based on narrow selectivity lectin affinity chromatography
Kwanyoung Jung
Department of Chemistry, Purdue University, West Lafayette, Indiana 47907, USA
J Proteome Res 8:643-50. 2009Lectin affinity chromatography using concanavalin A (Con A), Helix pomatia agglutinin (HPA), Lycopersicon esculentum lectin (LEL), Aleuria aurantia lectin (AAL) and Lens culinaris agglutinin (LCA) was used to investigate the utility of ..A new method for purification of carbonic anhydrase isozymes by affinity chromatography
O Ozensoy
Balikesir University, Science and Art Faculty, Department of Chemistry Biochemistry Section, 10100 Balikesir, Turkey
Biochemistry (Mosc) 69:216-9. 2004..Human carbonic anhydrase isozymes (HCA I and HCA II) and bovine carbonic anhydrase (BCA) were purified in high yields from erythrocytes...Separation of enzymatically active and inactive prostate-specific antigen (PSA) by peptide affinity chromatography
Ping Wu
Department of Clinical Chemistry, Helsinki University Central Hospital, Helsinki, Finland
Prostate 58:345-53. 2004..We have previously identified novel PSA-binding peptides that enhance the enzymatic activity of PSA when produced as fusion proteins...Efficient enrichment of intact phosphorylated proteins by modified immobilized metal-affinity chromatography
Anna Dubrovska
Ludwig Institute for Cancer Research, Biomedical Center, Uppsala University, SE 751 24 Uppsala, Sweden
Proteomics 5:4678-83. 2005..Here we describe an immobilized metal-affinity chromatography (IMAC)-based technique for the enrichment of phosphorylated proteins, which allows recovery of up to 90% ..Proteomic survey of copper-binding proteins in Arabidopsis roots by immobilized metal affinity chromatography and mass spectrometry
Cheng Che S Kung
Institute of BioAgricultural Sciences, Academia Sinica, Taipei, Taiwan, Republic of China
Proteomics 6:2746-58. 2006..systematically screened for copper-interacting proteins in Arabidopsis roots via copper-immobilized metal affinity chromatography (Cu-IMAC). We also compared Arabidopsis root metalloproteomes with affinity to Cu-IMAC and Zn-IMAC...Selective extraction and characterization of a histidine-phosphorylated peptide using immobilized copper(II) ion affinity chromatography and matrix-assisted laser desorption/ionization time-of-flight mass spectrometry
Scott Napper
Department of Biochemistry, Health Sciences Building, University of Saskatchewan, 107 Wiggins Road, Saskatoon, SK S7N 5E5 Canada
Anal Chem 75:1741-7. 2003..containing residues that form phosphoesters, such as serine and threonine, using immobilized metal-ion affinity chromatography. Despite the importance of histidine phosphorylation in two-component signal transduction pathways, ..Phosphorylation of the cytoplasmic domain of the integrin CD18 chain by protein kinase C isoforms in leukocytes
Susanna Fagerholm
MRC Protein Phosphorylation Unit, MSI WTB complex, University of Dundee, Dow Street, Dundee DD1 5EH, Scotland, United Kingdom
J Biol Chem 277:1728-38. 2002..When leukocyte extracts were subjected to affinity chromatography on agarose to which residues 751-761 of the CD18 chain phosphorylated at Thr-758 were bound covalently, ..Screening of protein-ligand interactions by affinity chromatography
Carlos D Garcia
Department of Chemistry, 118 Hand Chemical Laboratory, Mississippi State University, Mississippi State, Mississippi 39762, USA
Biotechnol Prog 19:575-9. 2003This paper examines affinity chromatography (AC) as an alternative tool for the determination of protein-ligand interactions for the particular case in which the ligand is the same protein...Determination of binding constants by affinity chromatography
Donald J Winzor
Department of Biochemistry, School of Molecular and Microbial Sciences, University of Queensland, Brisbane, QLD 4072, Australia
J Chromatogr A 1037:351-67. 2004This review summarizes developments in the use of affinity chromatography to characterize biospecific interactions in terms of reaction stoichiometry and equilibrium constant...Inactivation of herpes simplex type 1 gene vector on immobilized metal affinity chromatography: oxidative damage by hydroxyl free radicals and its prevention
Canping Jiang
Department of Chemical Engineering, University of Pittsburgh, Pittsburgh, Pennsylvania 15261, USA
Biotechnol Bioeng 95:48-57. 2006..The occurrence of MCO in immobilized metal affinity chromatography (IMAC) systems and its potential for inactivating biological products has not been well recognized...Using affinity chromatography to investigate novel protein-protein interactions in an undergraduate cell and molecular biology lab course
Kenneth D Belanger
Department of Biology, Colgate University, Hamilton, NY 13346, USA
CBE Life Sci Educ 8:214-25. 2009..Using recombinant protein expression, protein extraction, affinity chromatography, SDS-polyacrylamide gel electrophoresis, and Western blotting, undergraduates in a sophomore-level lab ..Isolation and biochemical characterization of highly purified Escherichia coli molecular chaperone Cpn60 (GroEL) by affinity chromatography and urea-induced monomerization
A Blennow
Department of Biochemistry, Chemical Center, University of Lund, Sweden
Biochim Biophys Acta 1252:69-78. 1995..purified from tightly bound substrate polypeptides by two different procedures: (i) group-specific affinity chromatography by using the triazine dye Procion yellow HE-3G as affinity ligand, and (ii) urea-induced monomerization ..
Research Grants
- Cardiotrophin-Like Cytokine 1, a Candidate Molecule for the FSGS FactorVirginia J Savin; Fiscal Year: 2010..These techniques include gel chromatography to determine the protein size and electrical charge, affinity chromatography to determine binding properties, and mass spectrometry...
- STRUCTURE AND FUNCTION OF HEPARIN COFACTOR IIDouglas M Tollefsen; Fiscal Year: 2010..Size-fractionated DS oligosaccharides will be subjected to HCII affinity chromatography, and the structures of selected high- and low-affinity oligosaccharides will be determined...
- HORMONAL REGULATION OF MESSENGER RNA STABILITYDaniel R Schoenberg; Fiscal Year: 2013..Aim 1 will use tandem affinity chromatography to recover the Complex I mRNP, identify its constituent proteins, and determine their role in mRNP ..
- MAP KINASE IN ISLET FUNCTIONMelanie H Cobb; Fiscal Year: 2013..We will evaluate candidates that may mediate the actions of these drugs identified by drug-affinity chromatography. Finally, we will continue to investigate ERK1/2-dependent signaling events that control insulin gene ..
- Hydroxynonenal induces glutathione synthesis through JNKHenry Jay Forman; Fiscal Year: 2010..In Aim 1, we will identify potential TFs by DNA affinity chromatography and LC-MS/MS analysis and then use chromatin immunoprecipitation (ChiP) assays to identify the EpRE and ..
- Clostridium difficile Toxin Gene RegulationABRAHAM LINCOLN SONENSHEIN; Fiscal Year: 2010..In addition, general, unbiased searches for the relevant regulatory proteins will be carried out based on affinity chromatography. A fourth protein, TcdC, will be tested for its potential activity as an antagonist of TxeR...
- Probing localiztion, interactions, and effector properties of Shigella lpaDNICHOLAS EUGENE DICKENSON; Fiscal Year: 2010..Specifically, fluorescence co-localization, FRET, and affinity chromatography experiments will be utilized in order to both map the intracellular distribution of IpaD and identify ..
- The Role of IgE in Autoimmune Skin DiseasesJanet A Fairley; Fiscal Year: 2013..IgE autoantibodies will be isolated from the sera of BP patients and controls by multi-step affinity chromatography. BP IgE will be tested for its ability to trigger mast cell degranulation in the presence of antigen ..
- Monolithic Media and Technology for BioprocessingALEX G BONNER; Fiscal Year: 2011..commercial products are (a) devices for anion and cation exchange chromatography, (b) devices for affinity chromatography, (c) low cost, disposable devices for large scale capture and release chromatography, and (d) application ..
- GE Healthcare Fast Protein Liquid Chromatography FPLC product number 13470501.Jasti S Rao; Fiscal Year: 2010..Chromatography is commonly used in the purification of biological materials. Specifically, affinity chromatography involves the use of packing which has been chemically modified by attaching a compound with a specific ..
- Engineering a packaging nanomotor for delivery of RNA and other moleculesVenigalla B Rao; Fiscal Year: 2010..The his-tagged mutants will be overexpressed in E.coli and purified by affinity chromatography using 96-well Ni-Sepharose plates...
- Identifying Host-Factors Required for Toxin Entry and ToxicityERIK HETT; Fiscal Year: 2010..Alternatively, the best compounds with no known target will be applied to affinity chromatography to identify their host targets, which will help determine their mechanisms of action...
- TYPE II 3BETA HSD GENE--REGULATION OF DHEAS SYNTHESISPeter Hornsby; Fiscal Year: 2009..The bovine adrenal cortex will be used as a source of the protein which will be purified by affinity chromatography;To investigate the possible regulation of ZR cell number by apoptosis...
- PROTEIN SEQUENCING BY TANDEM MASS SPECTROMETRYDonald F Hunt; Fiscal Year: 2013..OGlcNAcylated proteins so that they can be enriched from complex sample mixtures by immobilized metal affinity chromatography. In addition to the above work, we will conduct experiments to define the posttranslational modifications ..
- Novel transgenic tools for analysis of 5HT2C receptor expression and functionRonald B Emeson; Fiscal Year: 2013..in behavior, receptor expression/function and the ability to purify the tagged receptor using tandem affinity chromatography. It is anticipated that the development of this novel transgenic strategy will not only provide tools to ..
- Thrombomodulin Receptors on Immune CellsJohn Morser; Fiscal Year: 2013..We will use the TM lectin domain-Fc fusion protein to isolate that receptor by affinity chromatography from Jurkat cells and identify it by proteomics...
- The Role of Bovine Sperm Acrosomal Membrane-Matrix Complex in FertilizationSubir K Nagdas; Fiscal Year: 2013..b>Affinity chromatography and immunoprecipitation will be utilized to identify protein(s) responsible for membrane-matrix adhesion...
- Junctional exocytosis and breakdown of the intestinal barrier in inflammationANDREI IVANOVICH IVANOV; Fiscal Year: 2013..fusion machinery will be analyzed by a combination of biochemical (biotinylation, immunoprecipitation, affinity chromatography), immunocytochemical and genetic (siRNA knock-down of SNARE proteins, overexpression of NSF and 1SNAP ..
- Cellular Mapping of Small Molecule/Nucleic Acid InteractionsMicah J Bodner; Fiscal Year: 2012..or biotinyl moieties that will allow the molecule to be selectively purified from cellular debris with affinity chromatography. The analogs will be crosslinked to DNA (RNA) using ultraviolet light, and the small molecule/nucleic ..
- Analysis of Candida albicans filamentation using SILACDEREK PAUL THOMAS; Fiscal Year: 2012..Stable Isotope Labeling with Amino acids in Cell culture (SILAC) in conjunction with Immobilized Metal Affinity Chromatography (IMAC) enrichment of phosphoproteins followed by mass spectrometric analysis...
- Ah receptor transcriptional regulation through a novel DNA binding siteSHELLY RENEE WILSON; Fiscal Year: 2012..Specific Aim 1will employ DNA-affinity chromatography using the NC-XRE binding site characterized by EMSA and functional studies to fractionate the complex ..
- The molecular bases of inherited urea cycle disorders and ureagenesis regulationMendel Tuchman; Fiscal Year: 2010..as well as the variable domain peptide (VDP) will be characterized by immunoprecipitation, protein affinity chromatography, analytical ultracentrifugation, protein cross-linking and mass spectrometry, and the effects on ..
- Delivery of passive immunity against HIV using commensal vaginal Lactobacillus biLaurel A Lagenaur; Fiscal Year: 2012..The secreted antibodies in culture media will be purified using affinity chromatography and tested for neutralization of primary HIV-1 isolates in cell line-based and PBMC-based assays...
- Carcinogenic Metabolites Formed from AntiestrogenJudy L Bolton; Fiscal Year: 2013..The targeted proteins will be isolated using avidin affinity chromatography, separated by 2D electrophoresis, digested, and analyzed by MALDI-TOF and LC-MS-MS...
- Mechanistic Studies on Lung Cancer Prevention by Tea PolyphenolsChung S Yang; Fiscal Year: 2012..To identify direct molecular targets for EGCG, we will use affinity chromatography and mass spectrometry in collaboration with Dr. Zigang Dong at the University of Minnesota...
- BIOCHEMICAL PATHWAYS OF 12-HETE AND 12-KETE METABOLISMJoshua Rokach; Fiscal Year: 2010..This includes the synthesis of 5- HETE affinity chromatography ligands for 5-HEDH purification, followed by sequencing and cloning of the enzyme...
- Mechanism of SPARC peptide FSEC inhibition of angiogenesis in neuroblastomaChuan He; Fiscal Year: 2013..with a diazirine motif with human neuroblastoma tumor tissue proteins;2) purifying binding partners by affinity chromatography, and 3) identifying protein partners with mass-spectrometry...
- Protein Therapy for Acute Lung InjuryJahar Bhattacharya; Fiscal Year: 2010..The expressed proteins will be purified from the cell lysates using Immobilized Metal Affinity Chromatography (IMAC)...
- NuMA function in nonmalignant and malignant breast cellsSOPHIE A LELIEVRE; Fiscal Year: 2012..by assessing the interaction of NuMA with components of SNF2h and BAF types of ATP-dependent CRCs by affinity chromatography and co- immunoprecipitation, and assessing the effect of inhibiting NuMA expression on the assembly and ..
- Identification of Tyrosine-Sulfated Proteins in the Male Genital TractKevin L Moore; Fiscal Year: 2012..Tyrosine-sulfated proteins will then be purified from wild type mice by PSG2 affinity chromatography and identified by microsequencing...
- PS1 and BACE1 regulate processing and activity of voltage-gated sodium channelsDora M Kovacs; Fiscal Year: 2012..Cytoplasmic and nuclear binding partners of P2-ICD will be identified by immunoprecipitation and P2-ICD affinity chromatography. To characterize effects of p-/y-secretase-mediated processing of P2 on gene expression, gene expression ..
- Protein Arginine Methylation in TrypanosomesLAURIE READ; Fiscal Year: 2009..will be identified by anti-methylarg immunoprecipitation/mass spectrometry, and by TbPRMT6-PTP affinity chromatography. In Aim 3, we will analyze the mechanisms by which arg methylation impacts mitochondrial gene expression...
- Small molecule protein-glycan inhib. as malaria transmission-blocking therapueticRHOEL DAVID RAMOS DINGLASAN; Fiscal Year: 2013..glycosaminoglycan interactions involves (1) identifying novel lectin-like ookinete molecules by glycan-affinity chromatography and mass spectrometry, (2) characterizing their functional role in vivo through the production of gene ..
- Regulation of APP Pathway Gene Promoters in Alzheimer'sDebomoy K Lahiri; Fiscal Year: 2011..activity and ii) the status of such TF in normal and AD brain tissues using gel shift assay and DNA-affinity chromatography. 4) To characterize APP gene polymorphisms that influences the risk of late-onset Alzheimer's ..
- Functional characterization of Orientia tsutsugamushi ankryin repeat proteinsJason A Carlyon; Fiscal Year: 2013..and identify host cell target proteins that interact with Orientia Anks using in vivo coprecipitation ad affinity chromatography. The proposed work will fill a considerable knowledge gap of how an understudied pathogen of global ..
- MicroRNA networks in synaptic plasticityRichard H Goodman; Fiscal Year: 2010..cortical cells and transgenic mice, and complexes containing the associated microRNAs will be purified by affinity chromatography and identified by multiplex PCR...
- NMDA receptors in the diagnosis and treatment of pancreatic cancerWilliam G North; Fiscal Year: 2013..b>Affinity chromatography, IHC of human tissues, RIA, ELISA, RT-PCR, cloning, and DNA sequencing, Western analysis, and pathology ..
- Calcineurin Inhibition to Halt Aspergillus fumigatus Hyphae and VirulenceWilliam J Steinbach; Fiscal Year: 2010..We will purify and tag calcineurin pathway proteins and characterize their binding to effectors by affinity chromatography and differential dephosphorylation to elucidate interactions controlling the mechanism of calcineurin&..
- POLYNUCLEOTIDE HIGH PERFORMANCE AFFINITY CHROMATOGRAPHYHarry W Jarrett; Fiscal Year: 2013DESCRIPTION (provided by applicant): The goal is to improve the performance of the DNA-affinity chromatography of transcription factors and to increase understanding of genetic regulation. Many of the transcription factors (TFs;e.g...
- BIO-ORGANIC BIOMEDICAL MASS SPECTROMETRY RESOURCEAlma L Burlingame; Fiscal Year: 2013..Particularly in the last few years the tools of chromatography, affinity chromatography and advanced ion optical systems are poised with unprecedented analytical power to address ..
- BIO-ORGANIC BIOMEDICAL MASS SPECTROMETRY RESOURCEAlma L Burlingame; Fiscal Year: 2011..Particularly in the last few years the tools of chromatography, affinity chromatography and advanced ion optical systems are poised with unprecedented analytical power to address ..
- Nuclear Membrane Fusion in Xenopus Egg ExtractsMartin W Hetzer; Fiscal Year: 2010..We will use biochemical fractionation methods and affinity chromatography to identify the binding partner(s) of Importin b. 2...
- Protein Kinases of a TrypanosomeKojo Mensa Wilmot; Fiscal Year: 2010..In Specific Aim 2, we will validate the target of the protein kinase inhibitors, using a combination of affinity chromatography/chemical proteomics and molecular genetics approaches...
- Signature Peptide Approach to Biomonitor MDI ExposureADAM WISNEWSKI; Fiscal Year: 2013..Develop quantitative assay for measuring MDI-peptide purified from human serum samples by affinity chromatography, followed by LC-MS/MS. Aim 2...
- Glycobiological Analysis of Plasmodium-Vector Host InteractionsRHOEL DAVID RAMOS DINGLASAN; Fiscal Year: 2010..We also provide evidence for the use of lectin-affinity chromatography followed by tandem mass spectrometry to identify glycoproteins that are recognized by specific lectins...
- Mass Spectrometry Tools in Pursuit of Salivary Biomarkers of Sjogren's SyndromeSteven C Hall; Fiscal Year: 2013..In targeted approaches, we will use lectin and antibody affinity chromatography at the glycopeptide level with MS-based sequencing to discover pSS-associated differences in the ..
- Fusing Glycobiology and Nutritional Sciences to Prevent Necrotizing EnterocolitisLars Bode; Fiscal Year: 2009..AIM 2 will use Frontal Affinity Chromatography Mass Spectrometry to identify individual milk oligosaccharides with high affinity for P-selectin and ..
- Translational Control of Retroviral Unspliced mRNAKATHLEEN A BORIS LAWRIE; Fiscal Year: 2010..Combined results of site-directed mutagenesis, RNA affinity chromatography and MALDI-TOF mass spectroscopy have determined redundant stem-loop motifs are necessary for PCE activity ..
- Solubilization, purification, and crystallization of memebrane-associated proteinDavid G Myszka; Fiscal Year: 2010..receptors, 2) identifying conformationally sensitive antibodies for co- crystalization, 3) developing affinity chromatography protocols, 4) evaluating the effects of crystallization conditions on receptor activity, and 5) resolving ..
- The Role of Rab GTPases in Chlamydia-Infected CellsMarci A Scidmore; Fiscal Year: 2012..We will pursue yeast two-hybrid assays and affinity chromatography approaches to identify the chlamydial proteins that function to recruit Rab GTPases to the inclusion...
- Mechanism and function of ubiquitin binding by conserved endocytic protein Rvs167MARGARET MYERS; Fiscal Year: 2011..Interacting proteins will be identified by a whole genome yeast-2-hybrid screen, GST fusion protein affinity chromatography, or by coimmunoprecipitation from yeast lysates...
- Characterization of Innate immune receptorsDetlef Schuppan; Fiscal Year: 2009..innate immune receptor(s) on these cells that trigger(s) the innate immune response to gliadin by use of affinity chromatography of labeled cell membranes on the identified immobilized stimulatory gliadin peptide(s) and MALDI-TOF mass ..
- Advancement of Novel Small Molecules for Treatment of RabiesVishwanath R Lingappa; Fiscal Year: 2013..The advanced analogs have also been used as ligands for affinity chromatography to identify the host targets...
- G-Quadruplex-Mediated Transcriptional Regulation of PDGFR-??Danzhou Yang; Fiscal Year: 2013..For specific aim 3, we will use affinity chromatography and small molecule screening methods to identify proteins and drug-like molecules that bind to the ..
- Revealing Functions for Newly Discovered Proteins by FAST-NMRRobert Powers; Fiscal Year: 2010..Additionally, a high-performance affinity chromatography-mass spectrometry (HPAC-MS) component will be incorporated into the FAST-NMR screen to improve the ..
- Chemical probes of vulnerable pathways in antibiotic-resistant pathogensTanya Parish; Fiscal Year: 2013..2) We will use three methods (affinity chromatography, photoaffinity labeling, and yeast three hybrid system) to identify protein targets which bind to the ..
- Selection and Characterization of PSMA Ligands from Phage-Displayed LibrariesGary Fujii; Fiscal Year: 2006..Each anti-PSMA RRP fusion protein will be expressed in a bacterial system and purified by affinity chromatography. Anti-PSMA RRPs will then be tested in vitro for binding to purified PSMA ectodomain using qualitative ..
- Membrane Protein Co- Crystallization with Highly Crystalline and Soluble ProteinsGregory A Weiss; Fiscal Year: 2013..b>Affinity chromatography with the binding partner is aimed at assisting membrane protein purification, and co-crystallization aims ..
- Manufacturing of Trojan Horse-TNFR Decoy Receptor Fusion ProteinRuben J Boado; Fiscal Year: 2013..cultured in a 50L bioreactor, and the fusion protein will be purified with 1 liter columns or protein A affinity chromatography, cation exchange chromatography, and anion exchange chromatography...
- Thwarting HIV evasion of antibody avidity with novel antibody architecturesPamela J Bjorkman; Fiscal Year: 2012..Pooled bispecific reagents will separated by affinity chromatography to isolate the tightest binding pairs, which will be PCR amplified/sequenced to determine the two protein ..
- Thwarting HIV evasion of antibody avidity with novel antibody architecturesPamela J Bjorkman; Fiscal Year: 2013..Pooled bispecific reagents will separated by affinity chromatography to isolate the tightest binding pairs, which will be PCR amplified/sequenced to determine the two protein ..
- NEURAL NETWORK MODELING OF SPEECH PRODUCTIONFrank Guenther; Fiscal Year: 2000..P. gingivalis outer membrane proteins involved in hemin binding will be identified by hemin affinity chromatography. The specificity of the putative receptor(s) will be defined by examining the binding of 14[C]hemin to P...
- Genetics and Biochemistry of Sialylation in DrosophilaVladislav Panin; Fiscal Year: 2009..To this end, the sialyltransferase protein will be expressed in cell culture, purified using affinity chromatography, and assayed for its enzymatic activity...
- 14 3 3 Implications in Topoisomerase II PharmacologyDavid Kroll; Fiscal Year: 2003..Using reciprocal affinity chromatography and co-immunoprecipitation methods, we have shown that 14-3-3e, but NOT the G2 arrest protein 14-3-3E, ..
- CHEMISTRY AND METABOLISM OF SPHINGOLIPIDS & GLYCOPROTEINCHARLES SWEELEY; Fiscal Year: 1991..These enzymes will be purified from rat liver Golgi vesicles by affinity chromatography and their kinetic characteristics determined...
- Monoclonal Antibodies Against T-Regulatory CellsAlan Epstein; Fiscal Year: 2005..human lgG1 to create a fusion protein that can be produced in quantity and purified simply with Protein-A affinity chromatography. Purified human CD25-Fc will then be used in panning experiments using the MRC phage display library to ..
- CHARACTERIZATION OF THE PEMPHIGUS FOLIACEUS ANTIGENGrant Anhalt; Fiscal Year: 1993..treatment of viable epidermal cells, and the investigators could partially purify it by concanavalin A affinity chromatography and immunoprecipitation...
- BIOSPECIFIC POLYMER ENZYME CONJUGATES FOR DRUG DELIVERYGLEN KWON; Fiscal Year: 2004..A biotin moiety may mediate the separation of PEG-CPA conjugate by affinity chromatography, fractionating in terms of number of attached biotinylated PEG on CPA using an immobilized monomeric ..
- Novel Simulated Moving Bed Chromatography Device to Purify Recombinant Proteins ROBERT MIERENDORF; Fiscal Year: 2007..SMB) approach to the multi- milligram scale purification of recombinant proteins by immobilized metal affinity chromatography (IMAC)...