Histone

Histone H3K4me2 Antibody, SNAP-ChIP Certified

$465.00
SKU: 13-0027
Pack Size: 100 μg
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Type:  Monoclonal Host:  Rabbit
Mol Wgt.:  15 kDa Appl.:  ChIP, ChIP-Seq, IF, ICC, WB
Format:  Aff. Pur. IgG Reactivity: H, M, WR
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.

Histone H3K4me2 Antibody - SNAP ChIP Certified Application Notes Recommended dilutions:
 ChIP: 2 - 5 μg per 1x106 cells IF/ICC: 1 - 2 μg/mL WB: 0.02 - 0.2 μg/mL
Background References:
Grzybowski A et al. 2015 Mol Cell 58: 886-899 PMID 26004229

Product References:
Shah RN et al. 2018 A et al. 2015 Mol Cell 72: 162 PMID 30244833

View technical datasheet for this product. 13-0027 Datasheet

Applications Key: ChIP-Chromatin IP; E-ELISA; FACS-Flow cytometry; IF-Immunofluorescence; IHC-Immunohistochemistry; IP-Immunoprecipitation; WB-Western Blotting

Reactivity Key: B-Bovine; Ce-C. elegans; Ch-Chicken; Dm- Drosophila; Eu-Eukaryote; H-Human; M-Mouse; Ma-Mammal; R-Rat; Sc-S.cerevesiae; Sp-S. pombe; WR-Wide Range (predicted); X-Xenopus; Z-Zebrafish

13-0027 SNAP-ChIP Data

Representative SNAP-ChIP-Seq Results: Cumulative histogram plot and heatmap of signal intensity depict H3K4me2 ChIP-seq data aligned to annotated transcription start sites (TSS, +/- 3.0 kb; left). Two representative genomic regions depicting H3K4me2 peak structure and distribution are shown (right). Data shown are representative of H3K4me2 ChIP antibody (EpiCypher Catalog No. 13-0027) and are not lot-specific. Native ChIP-seq was performed using K562 cells as described (Shah et al., Mol Cell 2018) with SNAP-ChIPTM K-MetStat Spike-in (Catalog No. 19-1001) nucleosome controls added prior to chromatin digestion to confirm antibody specificity and ChIP efficiency. Paired-end sequencing libraries were prepared using the NEBNext® UltraTM II DNA Library Prep Kit for Illumina®. ChIP libraries were sequenced on an Illumina® NextSeq. Sequencing reads were aligned to the human genome using Bowtie 2 (Johns Hopkins University). Bigwig files of read enrichment in binned genomic regions (signal intensity) flanking the indicated gene features were used to create a cumulative histogram plot and heatmap of signal intensity ( www.basepairtech.com). Gene browser shots were generated using the Integrative Genomics Viewer (IGV, Broad Institute) with the window size denoted (top).
(Click image to enlarge)

13-0027 SNAP-ChIP Data

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)

13-0027 SNAP-ChIP Data

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)

13-0027 Western Blot data

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)

13-0027 Immunocytochemistry

Immunocytochemistry:ICC of HeLa cells, using Histone H3K4me2 Antibody (red). Actin filaments have been labeled with fluorescein phalloidin (green).
(Click image to enlarge)

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