Unit

UNIT 8.7 Isolation of Proteins for Microsequence Analysis

  1. Malcolm Moos Jr.

Published Online: 1 MAY 2001

DOI: 10.1002/0471142735.im0807s04

Current Protocols in Immunology

Current Protocols in Immunology

How to Cite

Moos, M. 2001. Isolation of Proteins for Microsequence Analysis. Current Protocols in Immunology. 8:III:8.7.

Author Information

  1. Center for Biologics Evaluation & Research Food and Drug Administration, Bethesda, Maryland

Publication History

  1. Published Online: 1 MAY 2001
  2. Published Print: DEC 1992

This is not the most recent version of the article. View current version (1 FEB 2007)

Abstract

The first basic protocol in this unit can be used to determine the amino-terminal sequence of polypeptides or proteins. It is particularly appropriate for large fragments of insoluble or hydrophobic proteins or proteins that cannot be purified to >90% molar purity without electrophoresis. Although the efficacy of this technique varies with the protein, it is possible to obtain useful sequence information starting with less than 50 pmol of the protein of interest. If the protein is blocked at the amino terminus, chemical cleavage or partial enzymatic digestion must be performed prior to electrophoresis. Upon isolation, the internal amino acid sequence is analyzed as described in the second basic protocol. This method requires ˜200 pmol of protein for analysis.