Pierce™ Agarose 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

Chromatin immunoprecipitation (ChIP) analysis was performed in albumin-treated NRK-52E cells using the following rabbit antibodies against: methylcytosine-binding protein 2 (MeCP2, 5 μg; ab2828, Abcam, Cambridge, UK), methyl-binding domain 1 (MBD1, 5 μg; sc-10751, Santa Cruz Biotechnology), acetylated histone H3 lysine 9 (H3K9ac, 2 μg; ab4441, Abcam), trimethylated histone H3 lysine 4 (H3K4me3, 2 μg; ab8580, Abcam) and NF-κB-p65 (5 μg; SC-372X, Santa Cruz Biotechnology) followed by qPCR using primers (ESM Table 1) in spanning genomic regions surrounding the miR-184 gene.

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Manufacturer protocol

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

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