UNIT 8.7 Isolation of Proteins for Microsequence Analysis
Published Online: 1 MAY 2001
Copyright © 2003 by John Wiley and Sons, Inc.
Lab Protocol Title
Current Protocols in Immunology
How to Cite
Moos, M. 2001. Isolation of Proteins for Microsequence Analysis. Current Protocols in Immunology. 8:III:8.7.
- Published Online: 1 MAY 2001
- Published Print: DEC 1992
This is not the most recent version of the article. View current version (1 FEB 2007)
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.