SNAP-ChIP K-MetStat Panel

Product Description

SNAP-ChIP (Sample Normalization and Antibody Profiling for Chromatin Immunoprecipitation) is a groundbreaking new technology that allows you to normalize your ChIP and ChIP-seq experiments as well as profile the specificity of your ChIP antibodies. SNAP-ChIP uses DNA barcoded recombinant designer nucleosomes (dNucs) as next-generation spike-ins for Chromatin Immunoprecipitation (ChIP). EpiCypher's K-MetStat panel can easily be added to any ChIP workflow without altering the protocol. However, users can monitor antibody specificity and evaluate technical variability within a ChIP or ChIP-seq experiment for the first time, setting SNAP-ChIP apart from any other exogenous spike-in controls (i.e. Drosophila) currently available, which do not assess antibody specificity or efficiency and are heterogenous & minimally reproducible.

The SNAP-ChIP K-MetStat Panel includes 1 unmodified nucleosome plus 15 modified nucleosomes with histone H3 or H4 posttranslational modifications (PTMs, created by a proprietary semi-synthetic method): H3K4me1, H3K4me2, H3K4me3, H3K9me1, H3K9me2, H3K9me3, H3K27me1, H3K27me2, H3K27me3, H3K36me1, H3K36me2, H3K36me3, H4K20me1, H4K20me2 and H4K20me3.

The panel consists of 16 pooled DNA-barcoded recombinant nucleosomes -

1 unmodified and 15 with lysine methyl marks on H3 and H4

SNAP-ChIP Data profiling antibody specificity:

Representative images for SNAP-ChIP K-MetStats (unmodified, H3K4me1, H3K4me2, H3K4me3) assayed in a chromatin immunoprecipitation (ChIP) experiment using commercially available ChIP grade antibodies (3 µg, n = 3). Quantitative Real-Time PCR (qPCR) for the DNA barcodes corresponding to unmodified (H3K4me0), H3K4me1, H3K4me2, and H3K4me3 nucleosomes show recovery of the barcodes corresponding to the expected antibody target. Identical experiments were performed for the remainder of the K-MetStat Panel (H3K9me1, H3K9me2, H3K9me3, H3K27me1, H3K27me2, H3K27me3, H3K36me1, H3K36me2, H3K36me3, H4K20me1, H4K20me2 and H4K20me3).

SNAP-ChIP is extremely useful for determining antibody cross-reactivity with off-target modifications. The cross-reactivity observed above (K4me1 & K4me2) is not indicative of a failure in the SNAP-ChIP procedure, but in the antibody to specifically recognize its target.