Anti-Histone H3 (di methyl K27) antibody - ChIP Grade (ab24684)

Protocol tips

Upstream tips	Protocol tips	Downstream tips
-Add protease inhibitors to all lysis solutions before use.	-Use 2-25 µg for µg of chromatin. -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.

Upstream tips
-Add protease inhibitors to all lysis solutions before use.
Protocol tips
-Use 2-25 µg for µg of chromatin. -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.

Publication protocol

Chromatin was extracted from naive CD4+Foxp3− T cells polarized for 48–72 h under various conditions (1×106 cells) after fixation with formaldehyde. Anti-H3K9me3 (61013; Active Motif), anti-dimethyl-H3 (diMe-Lys9) (D5567; Sigma-Aldrich), anti-H3K27me2 (ab24684; Abcam), anti-H3K4me3 (17-678; Millipore), anti-H3K36me3 (17-10493; Millipore), anti-H3K79me3 (17-10130; Millipore), anti-RORγt (sc-28559; Santa Cruz) and isotype-matched control antibody (sc-2027; Santa Cruz) were used for the immunoprecipitation of chromatin with an EZ ChIP kit according to the manufacturer’s instructions (Millipore). For cells transduced with retrovirus expressing FLAG-RelB, anti-FLAG (F1804; Sigma-Aldrich) was used. The precipitated DNA was then analyzed by real-time PCR. Data are presented as relative binding based on normalization to input DNA.

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Manufacturer protocol

Download the product protocol from Abcam for Anti-Histone H3 (di methyl K27) antibody - ChIP Grade (ab24684) below.

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