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Nuclear phosphorpteomics

Posted: Sun Jan 17, 2016 9:39 am
by ncmm
Hi All,
I want to perform nuclear phosphproteomics cancer cells (cell culture). Could some one suggest me a protocol for nuclear proteins extraction which can be further suitable for in-solution protein digestion?
thanks
ss

Re: Nuclear phosphorpteomics

Posted: Mon Jun 13, 2016 3:06 am
by Biomarker
Hey,

You can use https://tools.thermofisher.com/content/sfs/manuals/MAN0011398_NEPER_Nuc_Cytoplasmic_Extract_Reag_UG.pdf for nuclear protein extraction. I am not sure about the recepie of buffers but may be once lysis is done, you can do buffer exchange of lysate with 50 mM ammonium bicarbonate followed by in solution digestion. Hope it helps.