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Title Laboratory methods in enzymology : protein / edited by Jon Lorsch.
Published Amsterdam, Netherlands ; Boston, Mass. : Elsevier/Academic Press, 2014.
Book Cover

 Floor4  QP601 .M49 v.536 2014    IN LIBRARY
 Floor4  QP601 .M49 v.539 2014    IN LIBRARY
 Floor4  QP601 .M49 v.541 2014    IN LIBRARY
Description 3 v. : ill. ; 24 cm.
Series Methods in enzymology, 0076-6879 ; v. 536,539,541
Methods in enzymology ; v. 536.
Methods in enzymology ; v. 539.
Methods in enzymology ; v. 541.
Bibliography Includes bibliographical references and indexes.
Contents Part A. Practical steady-state enzyme kinetics -- Quantification of protein concentration using UV absorbance and Coomassie dyes -- Preparation of protein samples for mass spectrometry and N-terminal sequencing -- Lysis of mammalian and Sf9 cells -- In vivo (35 S)-methlionine incorporation -- Pulse-chase analysis to measure protein degradation -- Labeling of a protein with fluorophores using maleimide derivitization -- Labeling a protein with fluorophores using NHS ester derivitization -- Protein derivitization-expressed protein ligation -- Protein biotinylation -- Small-scale expression of proteins in E. coli -- Protein expression-yeast -- Recombinant protein expression in baculovirus-infected insect cells -- Single cell cloning of a stable mammalian cell line. Part B. In vitro synthesis of proteins in bacterial extracts -- Preparation of a Saccharomyces cervisiae cell-free extract for in vitro translation -- Yeast two-hybrid screen -- UV cross-linking of interacting RNA and protein in cultured cells -- Analysis of RNA-protein interactions by cell mixing -- General protein-protein cross-linking -- Chromatin immunoprecipitation and multiplex sequencing (ChIP-Seq) to identify global transcription factor binding sites in the nematode Caenorhabditis elegans -- PAR-CLIP (Photoactivatable ribonucleoside-enchanced crosslinking and immunoprecipitation): a step-by-step protocol to the transcriptome-wide identification of binding sites of RNA-binding proteins -- Determining the RNA specificity and targets of RNA-binding proteins using a three-hybrid system -- Dissecting a known RNA-protein interaction using a yeast three-hybrid system -- Identifying proteins that bind a known RNA sequence using the yeast three-hybrid system. Part C. TCA precipitation -- Coimmunoprecipitation of proteins from yeast -- Coupling antibody to cyanogen bromide-activated sepharose -- Analysis of protein-protein interactions by coimmunoprecipitation -- Use and application of hydrophobic interaction chromatography for protein purification -- Hydroxyapatite chromatography: purification strategies for recombinant proteins -- Salting out of proteins using ammonium sulfate precipitation -- Using ion exchange chromatography to purify a recombinantly expressed protein -- Gel filtration chromatography (size exclusion chromatography) of proteins -- Expression and purification of membrane proteins -- Explantory chapter: choosing the right detergent -- One-dimensional SDS-polyacrylamide gel electrophoresis (1D SDS-PAGE) -- Coomassie blue staining -- Silver staining of SDS-polyacrylamide gel -- Standard in vitro assays for protein-nucleic acid interactions-gel shift assays for RNA and DNA binding -- Protein filter binding -- Explanatory chapter: troubleshooting recombinant protein expression: general -- Explanatory chapter: troubleshooting protein expression: what to do when the protein is not soluble -- Western blotting using chemiluminescent substrates.
Subject Proteins -- Analysis.
Related Names Lorsch, Jon, editor of compilation.
ISBN 9780124200708
OCLC number 870866916