ChIP Rat - Brain microvessels

DNA-protein interactions are studied by using ChIP. The basic steps in this technique are crosslinking, sonication, immunoprecipitation, and analysis of the immunoprecipitated DNA. During ChIP, if chromatin is under-fragmented or fragments are too large which can lead to the increased background and lower resolution. Shorter cross-linking times (5-10 min) and/or lower formaldehyde concentrations (<1%) may improve shearing efficiency. If Chromatin is over-fragmented, then optimize shearing conditions for each cell type to improve ChIP efficiency. Over-sonication of chromatin may disrupt chromatin integrity and denature antibody epitopes. If you do not see any product or very little product in the input PCR reactions, add 5–10 μg chromatin per IP.

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Found 1 matching solution for this experiment

Imprint® Chromatin Immunoprecipitation Kit

Sigma-Aldrich

Upstream tips	Protocol tips	Downstream tips
	ChIP was performed using the Imprint Chromatin Immunoprecipitation Kit (Sigma-Aldrich) according to manufacturer’s instructions.

Protocol tips
ChIP was performed using the Imprint Chromatin Immunoprecipitation Kit (Sigma-Aldrich) according to manufacturer’s instructions.

Manufacturer protocol Publication protocol 1 Relevant paper

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