SimpleChIP® Plus Enzymatic Chromatin IP Kit (Magnetic Beads) #9005

Protocol tips

Upstream tips	Protocol tips	Downstream tips
-Once in solution, store 1M DTT at -20°C.	-For optimal ChIP results, use approximately 4 X 106 cells for each immunoprecipitation to be performed. -For optimal ChIP results, it is highly critical that the chromatin is of appropriate size and concentration. -For optimal ChIP results, use approximately 5 to 10 µg of digested, cross-linked chromatin per immunoprecipitation.

Upstream tips
-Once in solution, store 1M DTT at -20°C.
Protocol tips
-For optimal ChIP results, use approximately 4 X 106 cells for each immunoprecipitation to be performed. -For optimal ChIP results, it is highly critical that the chromatin is of appropriate size and concentration. -For optimal ChIP results, use approximately 5 to 10 µg of digested, cross-linked chromatin per immunoprecipitation.

Publication protocol

Chromatin immunoprecipitation was performed according to
previously published work. 2 Briefly, we used the SimpleChIP® Enzymatic Chromatin IP Kit (Cell
Signaling Technology) on isolated WT primary microglia grown to confluency in T75 flasks.
Following revers cross-linking, the samples were purified using the QIAquick PCR Purification
Kit (Qiagen). Quantitative PCR (qPCR) was performed using the SYBR qPCR master mix (Life
Technologies). Chromatin binding was calculated as a percentage of immunoprecipitated DNA
relative the input amount. Data were expressed as mean ± standard deviation of duplicate
samples from representative assays. The Rev-erb antibody used was developed in-house by
the Burris lab and has been described and validated previously3
. Good binding was determined
as compared to Gapdh binding.

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Manufacturer protocol

Download the product protocol from Cell Signaling Technology for SimpleChIP® Plus Enzymatic Chromatin IP Kit (Magnetic Beads) #9005 below.

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