ChIP-IT® Express Chromatin Immunoprecipitation Kits

Protocol tips

Upstream tips	Protocol tips	Downstream tips
-Do not re-freeze the Protein G magnetic beads, the beads should be stored at 4C.	-Always use ChIP validated antibody. -Optimal use of 1-3 ug of antibody.	-The diluted Proteinase K stop solution can not be stored.

Upstream tips
-Do not re-freeze the Protein G magnetic beads, the beads should be stored at 4C.
Protocol tips
-Always use ChIP validated antibody. -Optimal use of 1-3 ug of antibody.
Downstream tips
-The diluted Proteinase K stop solution can not be stored.

Publication protocol

Tissue fixation and chromatin isolation were carried out according to the Active Motif (Carlsbad, CA) protocol for ChIP. To validate the assay, Bhlhe40 antibody was used in a ChIP reaction with 19.5 μg of chromatin prepared from the cortex of a single female WT mouse (age 2.6 months). Chromatin was mixed with anti-Bhlhe40 antibody (Novus, cat# NB100-1800, Lot# C-1) and the ChIP DNA was processed into a standard Illumina ChIP-Seq library and sequenced. Antibody validation sequencing generated more than 5 million reads. Reads were aligned to the mouse genome (mm10), producing approximately 9.7 million unique alignments. MACS peak finding [24] was used to identify the most significant peaks using a default p value of 1 e-7, with 11,454 peaks identified. The antibody validation of Bhlhe40 ChIP-Seq indicated that the top motif identified by MEME [25] conformed to the CACGTG consensus E-box recognized by Bhlhe40.

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Manufacturer protocol

Download the product protocol from Active Motif for ChIP-IT® Express Chromatin Immunoprecipitation Kits below.

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