Anti-RUNX1 / AML1 antibody - ChIP Grade (ab23980)

Protocol tips

Upstream tips	Protocol tips	Downstream tips
-Add protease inhibitors to all lysis solutions before use.	-Use at an assay dependent concentration. -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 at an assay dependent concentration. -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

ChIP was performed using a SimpleChIP Plus Enzymatic Chromatin IP Kit (Cell Signaling Technology) according to the manufacturer’s instructions. In brief, cells were cross-linked in 1% formaldehyde in PBS for 10 minutes at room temperature. After glycine quenching, cell pellets were collected, lysed, and then sonicated with a Q55 sonicator system (QSONICA). The supernatant was diluted with the same sonication buffer, and processed for immunoprecipitation with the following antibodies at 4°C overnight: anti-RUNX1 (Abcam, ab23980), anti-RUNX2 (Cell Signaling Technology, D1L7F), and anti-RUNX3 (Abcam, ab11905). The beads were then washed and DNA was reverse cross-linked and purified. Following ChIP, DNA was quantified by qPCR using the standard procedures for the 7500 Real-Time PCR System.

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Manufacturer protocol

Download the product protocol from Abcam for Anti-RUNX1 / AML1 antibody - ChIP Grade (ab23980) below.

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