Pierce™ Magnetic ChIP Kit

Protocol tips

Upstream tips	Protocol tips	Downstream tips
If you are unfamiliar with cell type being used, culture an extra dish of cells for determining cell number. -Before crosslinking, trypsinize and determine the cell number from the extra dish of cells.	-After incubation, it is essential to open the column cap before plug removal to equalize the pressure within the column. Failure to open the cap before plug removal will result in sample loss.	-If you are preparing chromatin in bulk, unused supernatant may be stored at -80°C for later use.

Upstream tips
If you are unfamiliar with cell type being used, culture an extra dish of cells for determining cell number. -Before crosslinking, trypsinize and determine the cell number from the extra dish of cells.
Protocol tips
-After incubation, it is essential to open the column cap before plug removal to equalize the pressure within the column. Failure to open the cap before plug removal will result in sample loss.
Downstream tips
-If you are preparing chromatin in bulk, unused supernatant may be stored at -80°C for later use.

Publication protocol

A CHIP assay kit (Thermo‐Fisher Sci) was used as described previously (Liu et al. 2013). Briefly, cells were cross‐linked with 1.5% formaldehyde at 37°C for 10 min and collected in lysis buffer on ice. Cell lysates were sonicated and were diluted fivefold with a dilution buffer. Diluted cell lysates (500 μl) were used to immunoprecipitate protein‐DNA complexes. Immunoprecipitated DNA was amplified by PCR (30 cycles) using Taq polymerase (New England Biolabs) and then resolved by 1.2% agarose gel electrophoresis. The following primer set was used: forward, 5′‐CAC‐ATT‐TTT‐ATC‐TGG‐TCT‐AAT‐ACA‐CTG‐3′; and reverse, 5′‐ATT‐TTT‐AAT‐AGA‐ACT‐GAA‐AAG‐AGA‐AGT‐CA‐3′.

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Manufacturer protocol

Download the product protocol from Thermo Fisher Scientific for Pierce™ Magnetic ChIP Kit below.

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