Histone H3K27me3 Antibody, SNAP-ChIP Certified Description: This antibody meets EpiCypher’s “SNAP-ChIP® Certified” criteria for specificity and efficient target enrichment in a ChIP experiment (<20% cross-reactivity across the panel, >5% recovery of target input). Histone H3 is one of the four proteins that are present in the nucleosome, the basic repeating subunit of chromatin, consisting of 147 base pairs of DNA wrapped around an octamer of core histone proteins (H2A, H2B, H3 and H4). This antibody reacts to H3K27me3 and no cross reactivity
with H3K27me2, H3K27me1 or other lysine methylations in the EpiCypher SNAP-ChIP K-MetStat panel, is detected.
Histone H3K27me3 Antibody, SNAP-ChIP Certified Certified Immunogen: A synthetic peptide corresponding to histone H3 trimethylated at lysine 27.
Histone H3K27me3 Antibody, SNAP-ChIP Certified Certified Formulation: Protein A affinity-purified antibody (1 mg/mL) in PBS, with 0.09% sodium azide, 1% BSA, and 50% glycerol.
Histone H3K27me3 Antibody, SNAP-ChIP Certified Certified Storage and Stability:Stable for 1 years at -20°C from date of receipt.
ChIP qPCR Data: Histone H3K27me3 antibody (3 µg) was tested in a native ChIP experiment using chromatin from K-562 cells (3 µg) with the SNAP-ChIP K-MetStat Panel (EpiCypher Catalog No. 19-1001) spiked-in prior to micrococcal nuclease digestion. Specificity (left Y-axis) was determined by qPCR for the DNA barcodes corresponding to modified nucleosomes in the SNAP-ChIP panel (x-axis). Black bar represents antibody efficiency (right y-axis; log scale) and indicates percentage of the target immunoprecipitated relative to input. Error bars represent mean ± SEM in replicate ChIP experiments. (Click image to enlarge)
ChIP-seq Data: Histone H3K27me3 antibody (3 µg) was tested in a native ChIP experiment using chromatin from K-562 cells (3 µg) with the SNAP-ChIP K-MetStat Panel (EpiCypher Catalog No. 19-1001) spiked-in prior to micrococcal nuclease digestion. Ten nanograms ChIP DNA was subjected to library preparation using the NEBNext® UltraTM II DNA Library Prep Kit for Illumina®. ChIP libraries were analyzed by 2x150bp paired end sequencing on an Illumina HiSeq 4000. Paired reads were aligned to the SNAP-ChIP barcodes using the alignment algorithm available at https://www.basepairtech.com/. Specificity was determined by normalizing the read counts for each barcoded nucleosome IP to the corresponding Input and expressing the data as a percent of the nucleosome containing the target PTM (% Target). Error bars represent mean ± SEM for the duplicate DNA barcodes in a single ChIP experiment. (Click image to enlarge)
ChIP-seq Data: HChIP-seq was performed as described in Figure 2. Heatmap of H3K4me3 (left) compared to H3K27me3 (right) shows opposing patterns of enrichment (A). Representative gene loci for EpiCypher H3K27me3 antibody ChIP-seq tracks (green tracks) are compared to two different ENCODE tracks (blue), shows similar patterns of enrichment (Click image to enlarge)
Western Blot Data: Recombinant histone H3.3 (Lane 1) and acid extracts of HeLa cells (Lane 2) were blotted onto PVDF and probed with 1 µg/mL Histone H3K27me3 Antibody. (Click image to enlarge)
Immunohistochemistry Data: IHC staining of HepG2 cells using Histone H3K27me3 Antibody. (Click image to enlarge)