A Practical Guide to Protein and Peptide Purification for Microsequencing

✍ Scribed by Paul T. Matsudaira

• Publisher: Academic Press, Academic Press
• Year: 1993
• Tongue: English
• Leaves: 189
• Edition: 2
• Category: Library

✦ Synopsis

Why a Second Edition?
The Second Edition provides practical answers to the general question, "How can I obtain useful sequence information from my protein or peptide?" rather than the more specific question asked in the first edition, "How can I obtain the N-terminal sequence?" Important new methods include ways of dealing with blocked N termini, computer analysis of protein sequences, and the recent revolution in mass spectrometry.

Key Features
Highlights of the Second Edition:
Mass spectrophotometric characterization of proteins and peptides
N-terminal sequencing of proteins with blocked N termini
Internal amino acid sequence analysis after protease digestion in-gel and on-blot
Improved microscale peptide purification methods
Computer analysis of protein sequences
New protocols tested and refined through everyday use in authors' laboratories
* Updated reference chapter covering all aspects of protein microsequencing

✦ Table of Contents

Content:
Front Matter, Page iii
Copyright, Page iv
Contributors, Pages xi-xii
Preface, Pages xiii-xv, Paul Matsudaira
Preface to the First Edition, Pages xvii-xviii, Paul T. Matsudaira
Abbreviations, Pages xix-xx
Introduction, Pages 1-13, Paul Matsudaira
1 - Strategies for Obtaining Partial Amino Acid Sequence Data from Small Quantities (<500 pmol) of Pure or Partially Purified Protein, Pages 15-41, Harry Charbonneau
2 - Enzymatic Digestion of Proteins and HPLC Peptide Isolation, Pages 43-69, Kathryn L. Stone, Kenneth R. Williams
3 - Purification of Proteins and Peptides by SDS–PAGE, Pages 71-101, Nancy LeGendre, Michael Mansfield, Alan Weiss, Paul Matsudaira
4 - Internal Amino Acid Sequence Analysis of Proteins after in Situ Protease Digestion on Nitrocellulose, Pages 103-124, Ruedi Aebersold
5 - Mass Spectrometric Strategies for the Structural Characterization of Proteins, Pages 125-153, Hubert A. Scoble, James E. Vath, Wen Yu, Stephen A. Martin
Appendix - SEQUENCES OF COMMONLY USED PROTEASES, Pages 155-157
References, Pages 159-184