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Western blotting of histones from acid-urea-Triton- and sodium dodecyl sulfate-polyacrylamide gels.

Authors
Type
Published Article
Journal
Analytical biochemistry
Publication Date
Volume
162
Issue
2
Pages
430–434
Identifiers
PMID: 3605607
Source
Medline
License
Unknown

Abstract

We have developed a method for histone transfer from acid-urea-Triton (AUT)-polyacrylamide gels to nitrocellulose filters which prevents the interference of Triton X-100 with the binding of histones to nitrocellulose. Equilibration of AUT gels in 50 mM acetic acid and 0.5% sodium dodecyl sulfate (SDS) allowed displacement of Triton by SDS without loss of band resolution. Electrotransfer of all histone species from treated AUT gels or from equilibrated SDS gels was complete within 1 h in a transfer buffer of Tris-glycine with SDS for increased transfer efficiency and methanol for histone binding. Nitrocellulose with a pore size of 0.2 micron was optimal for histone detection.

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