Histone

Histone H3K4me3 Antibody, SNAP-ChIP Certified *DISCONTINUED*

SKU: 13-0028
Pack Size: 100 μg
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Product Discontinued - See 13-0060 as a recommended alternative

Type:  Monoclonal Host:  Rabbit
Mol Wgt.:  15 kDa Appl.:  ChIP, ChIP-Seq, ELISA
Format:  Aff. Pur. IgG Reactivity: H, M, WR
Histone H3K4me3 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 H3K4me3 and no cross reactivity with H3K4me1 or H3K4me2, or other lysine methylations in the EpiCypher SNAP-ChIP K-MetStat panel, is detected.
Histone H3K4me3 Antibody - SNAP ChIP Certified Immunogen: A synthetic peptide corresponding to histone H3 trimethylated at lysine 4.

Histone H3K4me3 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 H3K4me3 Antibody - SNAP ChIP Certified Storage and Stability: Stable for 1 years at -20°C from date of receipt.

Histone H3K4me3 Antibody - SNAP ChIP Certified Application Notes Recommended dilutions:
ChIP 2 - 5 μg per 1x106 cells ELISA 1 - 10 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
Lam K-WG et al. 2019 Nat. Commun. 10: 3821 PMID 31444359

View technical datasheet for this product. 13-0028 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

13-0028 SNAP-ChIP Data

Representative SNAP-ChIP-seq results: Cumulative histogram plot and heatmap of signal intensity depict H3K4me3 ChIP-seq data aligned to annotated transcription start sites (TSS, +/- 3.0 kb; left). Two representative genomic regions depicting H3K4me3 peak structure and distribution are shown (right). Data shown are representative of H3K4me3 ChIP antibody (EpiCypher Catalog No. 13-0028) 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)

13-0028 SNAP-ChIP Data

SNAP-ChIP qPCR Data: Histone H3K4me3 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)

13-0028 SNAP-ChIP Data

SNAP-ChIP-seq Data: Histone H3K4me3 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)

13-0028 ELISA

ELISA Data: Designer Nucleosomes (dNucs) containing the indicated H3K4 modification were used in an ELISA assay with Histone H3K4me3 Antibody (10 μg/mL).
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