I am using western blot to check status of acetylated H3 in adipose tissues (total homogenization by RIPA), but it does not work, maybe I have to extract histone protein. I hope someone has experience about this can give me some advice.
Dear Athanasakis,
Thank so much for responding for my question and giving me your acid extraction protocol. I am trying to do western blot with whole homogenization, because I would like to check other proteins as well in the sample which I want to see how is histone acetylation. I do not have enough samples to divide them, one for histone and another for other proteins.
Hope to hear other recommendation!
Hi Dinh-Toi, I've not used this kit but am also hoping to find a good way to prep protein samples so that I can look at histones as well as other proteins. http://www.piercenet.com/product/subcellular-protein-fractionation-kit-tissues Kit indicates that it will give a chromatin-bound fraction which should work well to look at histones and modified histones, as well as cytoplasmic, nuclear and a few other fractions.
Thank Dr. Stephanie for sharing me information of the kit, I have taken a look, and think that is a good stool to prepare protein samples for histones. But, fist at all, I will try to extract histone by Abcam' protocol, and RIPA buffer (without spinning) to see whether they are working or not.
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