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

The ChIP experiments were carried out as previously described [64,66]. The Magna ChIP kit was purchased from Thermo-Fisher Scientific (Rockford, IL). The chromatin samples were pre-cleared, then subjected to immunoprecipitation with anti-human OCT4 antibody bound to beads. The chromatin immunoprecipitate complexes were analyzed by PCR using primer pairs (For-5'-GGGAAATAGCGGGAATGTTG-3' and Rev-5'-GCGAAATGCCCAGAACTC-3'; NCBI reference sequence: NC_000004.12) that amplify an ~850-bp DNA sequence in the region of the human VEGFR-2/FLK1-promoter/enhancer. The PCR product was resolved by 4% agarose gel in 1× TAE buffer, thereafter visualized by staining with 0.2 μg/ml ethidium bromide (EtBr), and images captured using a Polaroid gel documentation system.

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Manufacturer protocol

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

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