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

✍ Scribed by Jakob H. Waterborg; Rodney E. Harrington

Publisher: Elsevier Science
Year: 1987
Tongue: English
Weight: 1023 KB
Volume: 162
Category: Article
ISSN: 0003-2697
DOI: 10.1016/0003-2697(87)90415-5

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✦ Synopsis

We have developed a method for histone transfer from acid-urea-Triton (AUT)-polyacrylamide gels to nitrocellulose titers which prevents the interference of Triton X-100 with the binding of histones to nitrocelhtlose. 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 pm was optimal for histone detection. o 1987 Academic mess hc.

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