Histone H3K4me2 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 H3K4me2 and no cross reactivity with H3K4me1 or H3K4me3, or other lysine methylations in the EpiCypher SNAP-ChIP K-MetStat panel, is detected.
Histone H3K4me2 Antibody - SNAP ChIP Certified Immunogen: A synthetic peptide corresponding to histone H3 dimethylated at lysine 4.
Histone H3K4me2 Antibody - SNAP ChIP Certified Formulation: Protein A affinity-purified antibody (1 mg/mL) in PBS, with 0.09% sodium azide, 1% BSA, and 50% glycerol.
Histone H3K4me2 Antibody - SNAP ChIP Certified Storage and Stability:Stable for 1 years at -20°C from date of receipt.
SNAP-ChIP qPCR Data:Histone H3K4me2 antibody (3 μg) was tested in a native ChIP experiment with chromatin from HEK-293 cells (~1x106 cells) 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)
SNAP-ChIP-seq Data: Histone H3K4me2 antibody (5 μg) was tested in a native ChIP experiment using chromatin from K-562 cells (~1x106 cells) with the SNAP-ChIP K-MetStat Panel (~1x106 cells)
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)
Western Blot Data: Recombinant histone H3.3 (Lane 1) and acid extracts of HeLa cells (Lane 2) were blotted onto PVDF and probed with 0.025 μg/mL Histone H3K4me2 Antibody. (Click image to enlarge)
Immunocytochemistry:ICC of HeLa cells, using Histone H3K4me2 Antibody (red). Actin filaments have been labeled with fluorescein phalloidin (green). (Click image to enlarge)