Hello everyone,
I wanted to detect histone protein expression levels in untreated and treated cell culture samples. Few research articles demonstrate histone can be isolated by usual lysis buffer and total protein lysate will be obtained in supernatant. On contrary few articles demonstrate that histone can be isolated by HCL or sulphuric acid followed by TCA precipitation method.
If someone suggests which is best method to isolate total histones would be helpful to me.
TIA
Sudheer
Boris Leibovitch
In total cell lysate you will get only "loose" proteins (and only some histones) while most of histone rather tightly bound to DNA and will be in precipitate after lysis and centrifugation. I would advise any of harsher methods, including HCl.
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