SNAP-ChIP - K-MetStat Panel
A panel of distinctly modified mononucleosomes assembled from recombinant human histones expressed in E. coli (two each of histones H2A, H2B, H3 and H4; accession numbers: H2A-P04908; H2B-O60814; H3.1-P68431 or H3.2-Q71DI3*; H4-P62805) wrapped by 147 base pairs of barcoded Widom 601 positioning sequence DNA. The panel is comprised of a pool of 1 unmodified plus 15 histone H3 or H4 post-translational 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. Each distinctly modified nucleosome is distinguishable by a unique sequence of DNA ("barcode") at the 3` end that can be deciphered by qPCR or next-generation sequencing. Each of the 16 nucleosomes in the pool is wrapped by 2 distinct DNA species, each containing a distinct barcode ("A" and "B", see SNAP-ChIP Manual) allowing for an internal technical replicate.
SNAP-ChIP K-MetStat Panel Formulation: Purified recombinant mononucleosomes, containing a mixture of 16 (1 unmodified plus 15 unique) H3 and H4 PTMs in 10 mM sodium cacodylate, pH 7.5, 100 mM NaCl, 1 mM EDTA, 50% glycerol (w/v), 1x Protease Inhibitor cocktail, 100 ug/mL BSA, 10 mM β-mercaptoethanol. Molarity = 0.6 nM. MW = ~199382.1 Da (average MW of all 16 nucleosomes).
SNAP-ChIP K-MetStat Panel Storage and Stability: Stable for six months at -20°C from date of receipt.
SNAP-ChIP K-MetStat Panel Application Notes: SNAP-ChIP K-MetStats are highly purified recombinant mononucleosomes and are suitable for use as spike-in controls for ChIP reactions, for antibody specificity testing or for effector protein binding experiments. SNAP-ChIP K-MetStats from EpiCypher do not contain detectable levels of free DNA which could alter assayed activities. See manual for more information. SNAP-ChIP can easily be introduced into existing ChIP and ChIP-seq workflows for experiment normalization and monitoring of technical variability.