MAGnify™ Chromatin Immunoprecipitation System

Protocol tips

Upstream tips	Protocol tips	Downstream tips
-The 1.25 M glycine must be at room temperature before use. -The Lysis Buffer must be at room temperature and fully resuspended before use. Vortex briefly to resuspend.	-For each ChIP reaction, use 10,000–300,000 cells or 0.167–5 mg of tissue. -1–10 μg of antibody is a typical starting range. - 10-minute crosslinking step using formaldehyde at a 1% final concentration. -Keep the cell lysate cooled on ice during sonication.

Upstream tips
-The 1.25 M glycine must be at room temperature before use. -The Lysis Buffer must be at room temperature and fully resuspended before use. Vortex briefly to resuspend.
Protocol tips
-For each ChIP reaction, use 10,000–300,000 cells or 0.167–5 mg of tissue. -1–10 μg of antibody is a typical starting range. - 10-minute crosslinking step using formaldehyde at a 1% final concentration. -Keep the cell lysate cooled on ice during sonication.

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 Thermo Fisher Scientific for MAGnify™ Chromatin Immunoprecipitation System below.

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