Histone extraction, and... - (Oct/11/2006 )
I've been through several protocols for histone extraction, but what I am looking for is the one which allows extraction histones without losing other nuclear proteins in process.
Is a simple method of adding DNAse to nuclear extracts going to help? I'd like to do one extraction for my blots, and not two separate ones for histones and other proteins.
Is acetylation and phosphorylation affected with the standard nuclei isolation protocol?
I'm not exactly interested in puryfying histones, but making an extract that I can use for acetylated, phosphorylated histones AND nuclear proteins.
It would be easier to answer your question if we knew
the specific experiment you were planning on doing.
Are you doing Westerns for modified histones?
However, I can say that histones can be isolated along with
other nuclear proteins if you soncate the DNA with low intensity pulses.
M____
the specific experiment you were planning on doing.
Are you doing Westerns for modified histones?
However, I can say that histones can be isolated along with
other nuclear proteins if you soncate the DNA with low intensity pulses.
M____
Yes, exactly that - westerns on modified histones, and for proteins that play along with the modifications (and to have them all in one sample is, of course, not only convenient, but makes the experimtent more accurate).
Ummm, sonication might be a good idea.
I'll most certainly try it to Telomerase and others:
I want to do the western blot to detect the histone methylation in rice (Oryza sativa ), could you send me the protocols for histone extraction in detail ?
my first time to do it, can you give me some suggestions ? thank you very much
qinfujun