Magna ChIP™ G Tissue Kit

Protocol tips

Upstream tips	Protocol tips	Downstream tips
- DO NOT cryo-preserve specimen with sucrose. -Make fresh 1% formaldehyde before each experiment.	-Keep lysate ice-cold. Sonication produces heat, which can denature the chromatin. Allow time between cycles of sonication to prevent sample overheating. - DNA fragments should be in 200 -1000 bps in size. -Add 1-10 µg of immunoprecipitating antibody.

Upstream tips
- DO NOT cryo-preserve specimen with sucrose. -Make fresh 1% formaldehyde before each experiment.
Protocol tips
-Keep lysate ice-cold. Sonication produces heat, which can denature the chromatin. Allow time between cycles of sonication to prevent sample overheating. - DNA fragments should be in 200 -1000 bps in size. -Add 1-10 µg of immunoprecipitating antibody.

Publication protocol

For ChIP, the MAGnify Kit (ThermoFisher) was used following the manufacturer’s recommendations.
2 x 106
primary adult murine CFs were seeded in a 10 cm petri dish per experimental condition. Crosslinking of cells was performed directly on the dish without trypsinization. Sonication was performed
on a Bioruptor UCD-200TM-EX for 16 cycles of 30 seconds ON, 30 seconds OFF at high power.
200.000 cells were used for each immunoprecipitation. For P53, 4 µg of a mouse monoclonal anti-P53
antibody (Cell Signaling 2524S) were used, whereas for IgG control IP, 4 µg mouse monoclonal
isotype control (Cell Signaling 5415S) were used.

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Manufacturer protocol

Download the product protocol from Merck Millipore for Magna ChIP™ G Tissue Kit below.

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