Lysates from each group were generated with the Piece Co-IP Kit (Thermo Scientific, Bonn, Germany). Then the co-immunoprecipitation (Co-IP) was carried out according to the manufacturer's instructions. Briefly, the rabbit polyclonal β-catenin antibody (Cell Signaling Technology, Danvers, MA, USA) was first immobilized for 2 h onto Amino Link Plus coupling resin. The resin was then washed and incubated with lysate overnight at 4°C. Subsequently, the resin was washed again and then eluted by elution buffer. Elution buffers were collected and analyzed by Western blotting which used rabbit polyclonal β-catenin antibody(Cell Signaling Technology, Danvers, MA,USA), rabbit polyclonal p300 antibody antibody (Santa Cruz Biotechnology), rabbit polyclonal acetylated-lysine antibody(Cell Signaling Technology) and peroxidase-conjugated goat anti-rabbit IgG (Bioss). A negative control was employed in the Co-IP kit to assess non-specific binding.
Ling F., Tang Y., Li M., Li Q.S., Li X., Yang L., Zhao W., Jin C.C., Zeng Z., Liu C., Wu C.F., Chen W.W., Lin X., Wang Y.L, & Threadgill M.D. (2017). Mono-ADP-ribosylation of histone 3 at arginine-117 promotes proliferation through its interaction with P300. Oncotarget, 8(42), 72773-72787.
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