Immunodetection after complete destaining of Coomassie blue-stained proteins on immobilon-PVDF

Jon L. Pryor, Wenhao Xu, David W. Hamilton

Research output: Contribution to journalArticlepeer-review

19 Scopus citations

Abstract

A technique that simplifies the localization of an immunodetectable protein in relation to the other electrophoresed proteins is described. Proteins are transblotted onto a polyvinylidene difluoride (PVDF) membrane and visualized by staining with Coomassie brilliant blue R-250, and a photograph of the protein pattern is taken. The Coomassie blue-stained PVDF membrane is then completely destained using a 25% acetic acid/50% methanol solution that allows subsequent immunostaining on the same membrane. The technique uses common laboratory reagents, is rapid, and has been shown to be applicable for a variety of proteins using both monoclonal and polyclonal antibodies and a variety of transblots.

Original languageEnglish (US)
Pages (from-to)100-104
Number of pages5
JournalAnalytical Biochemistry
Volume202
Issue number1
DOIs
StatePublished - Apr 1992

Bibliographical note

Funding Information:
USDA Grant Foundation

Funding Information:
by USPH Grant from the Andrew

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