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

ChIP analysis was performed using Chromatin IP kit (Pierce). Anti-p65(pNF-κB) antibody (CST 8242S) was used for immunoprecipitation and then pNF-κB association with GM-CSF promoters was estimated using SYBR-Green RT-PCR. For positive control polymerase II antibody was used (Cell Signaling Technology). Corresponding normal IgG was used as negative control. ChIP was performed overnight at 4 °C with rotation. Input DNA was purified along with ChIP probes. Association with pNF-κB and GM-CSF promoters was estimated using SYBR-Green RT-PCR.

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Manufacturer protocol

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

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