Laboratory Methods in Enzymology: Protein Part A

Edited by

  • Jon Lorsch, Department of Biophysics and Biophysical Chemistry, Johns Hopkins University School of Medicine, Baltimore, MD, USA

The critically acclaimed laboratory standard for almost 50 years, Methods in Enzymology is one of the most highly respected publications in the field of biochemistry. Each volume is eagerly awaited, frequently consulted, and praised by researchers and reviewers alike. Now with more than 530 volumes and 40,000 chapters in the collection, this is an essential publication for researchers in all fields of life sciences, including microbiology, biochemistry, cancer research, and genetics, just to name a few.

This volume brings together a number of core protocols concentrating on protein, carefully written and edited by experts, including:

  • Pulse-chase analysis to measure protein degradation
  • Labeling a protein with fluorophores using NHS ester derivitization
  • Immunoaffinity purification of proteins
  • Proteolytic affinity tag cleavage
  • Purification of GST-tagged proteins

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Audience

Biochemists, biophysicists, molecular biologists, analytical chemists, and physiologists

 

Book information

  • Published: January 2014
  • Imprint: ACADEMIC PRESS
  • ISBN: 978-0-12-420070-8


Table of Contents

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]-Methionine 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
Purification of His-tagged proteins
Affinity purification of a recombinant protein expressed as a fusion with the maltose-binding protein (MBP) tag
Immunoaffinity purification of proteins
Affinity Purification of Protein Complexes Using TAP Tags
Strep-tagged protein purification
Proteolytic affinity tag cleavage
Affinity Pull-down of Proteins using anti-FLAG M2 Agarose Beads
Protein Affinity Purification using Intein/Chitin Binding Protein Tags
Purification of GST-tagged proteins