Anti-Histone H3 (tri methyl K4) antibody - ChIP Grade (ab8580)

Protocol tips

Upstream tips	Protocol tips	Downstream tips
-Add protease inhibitors to all lysis solutions before use.	-Keep cells on ice between the rounds of homogenisations. - Increase or decrease the homogenization step to maximize the yield of nuclei depending on cell line. -Do not place ethidium bromide in the agarose gel or the electrophoresis buffer, because of the presence of SDS.

Upstream tips
-Add protease inhibitors to all lysis solutions before use.
Protocol tips
-Keep cells on ice between the rounds of homogenisations. - Increase or decrease the homogenization step to maximize the yield of nuclei depending on cell line. -Do not place ethidium bromide in the agarose gel or the electrophoresis buffer, because of the presence of SDS.

Publication protocol

For both cell line and tissue ChIPs, 5 μg of antibody and 50 μl of magnetic protein A or G beads (10008D or 10009D; Thermo Fisher Scientific) were used per IP. The following antibodies were used: ERα (HC-20, sc-543 Santa Cruz Biotechnology lot K1715 and F1215), Foxa1/2 (M-20, sc-6554 Santa Cruz Biotechnology lot G1415 and D1015), AR (N-20, sc-816 Santa Cruz Biotechnology lot C0916 and I0215), H3K27ac (39133Active Motif lot 31814008), and H3K4me3 (ab8580 Abcam lot GR240214-2).

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Manufacturer protocol

Download the product protocol from Abcam for Anti-Histone H3 (tri methyl K4) antibody - ChIP Grade (ab8580) below.

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