Rapid and effective western blotting of histones from acid-urea-Triton and sodium dodecyl sulfate polyacrylamide gels: two different approaches depending on the subsequent qualitative or quantitative analysis.
An improved method for the electrophoretic transfer of histones from sodium dodecyl sulfate (SDS) and acetic acid-urea-Triton X-100 (AUT) polyacrylamide gels onto nitrocellulose membranes is described. In the case of SDS-gels, it was not essential to equilibrate them before transfer while for the AUT-gels, an equilibration step is essential to prevent the interference of Triton X-100 with the binding of histones to nitrocellulose. Transfer efficiency was different for different histone classes. Two procedures were developed: (1) one suitable for qualitative studies, and (ii) another for quantitative transfer.
Thiriet
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C
Laboratoire de Biologie Cellulaire, Université de Reims, France.
Albert
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P
D000085:Acetates; D019342:Acetic Acid; D000818:Animals; D015153:Blotting, Western / Q000379:methods*; D002467:Cell Nucleus / Q000737:chemistry; D002645:Chickens; D003101:Collodion; D004591:Electrophoresis, Polyacrylamide Gel / Q000379:methods*; D004912:Erythrocytes / Q000648:ultrastructure; D006657:Histones / Q000302:isolation & purification*; D017830:Octoxynol; D012967:Sodium Dodecyl Sulfate; D014508:Urea
D000085:Acetates; D006657:Histones; D012967:Sodium Dodecyl Sulfate; D014508:Urea; D017830:Octoxynol; D003101:Collodion; D019342:Acetic Acid