I want to use isolated histones with the Abcam Histone H3 Modification Multiplex Assay Kit (ab185910) and need to know if using a trichloroacetic acid or acetone precipitation step when isolating the histones will preserve or destroy histone modifications such as acetylation, methylation, and phosphorylation for detection by antibodies.
Yamini Dalal
Instead of the kit, which uses either TCA or acetone (you can lose phosphorylation with acids), if you have enough cells, you could try hydroxyapatite purification, eluted by 2M NaCl, and concentrate the 2M eluates
using an Amicon. You can dialyze out the salt vs. 150mM NaCl-TE buffer (see Walkiewicz, Bui et al from my lab on the full protocol). Good luck, happy histone mod hunting! Cheers, Yamini
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