A simple in situ cyanogen bromide cleavage method to obtain internal amino acid sequence of proteins electroblotted to polyvinyldifluoride membranes

Mitchell G. Scott, Daniel L. Crimmins, David W. McCourt, Jeffrey J. Tarrand, Margaret C. Eyerman, Moon H. Nahm

Research output: Contribution to journalArticle

62 Scopus citations

Abstract

We report a simple method to obtain internal amino acid sequences from larger proteins electroblotted to polyvinyldifluoride membranes. To demonstrate this method, immunoglobulin heavy and light chains are separated by gel electrophoresis and electroblotted to the membrane. The separated chains, immobilized to the membrane, are cleaved in situ by cyanogen bromide and the resulting fragments are subsequently eluted from the membrane. The fragments are separated by gel electrophoresis, electroblotted and subjected to gas phase microsequence analysis.

Original languageEnglish
Pages (from-to)1353-1359
Number of pages7
JournalBiochemical and Biophysical Research Communications
Volume155
Issue number3
DOIs
StatePublished - Sep 30 1988
Externally publishedYes

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