Western blotting of histones from acid-urea-Triton- and sodium dodecyl sulfate-polyacrylamide gels.
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.
D000818:Animals; D002417:Cattle; D003101:Collodion; D004563:Electrochemistry; D004591:Electrophoresis, Polyacrylamide Gel / Q000379:methods; D006657:Histones / Q000302:isolation & purification*; D006863:Hydrogen-Ion Concentration; D017830:Octoxynol; D011092:Polyethylene Glycols; D012967:Sodium Dodecyl Sulfate; D013950:Thymus Gland / Q000032:analysis; D014508:Urea
D006657:Histones; D012967:Sodium Dodecyl Sulfate; D011092:Polyethylene Glycols; D014508:Urea; D017830:Octoxynol; D003101:Collodion