Histone H3K27ac Antibody, SNAP-ChIP® Certified

SKU: 13-0045
Pack size: 50 µg
Type:  Monoclonal Host:  Mouse
Mol Wgt.:  15 kDa Appl.:  ChIP, ChIP-Seq, L, IF
Format:  Aff. Pur. IgG Reactivity: H, M, WR
Histone H3K27ac 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 H3K27ac and no cross reactivity with other lysine acylations in the EpiCypher SNAP-ChIP K-AcylStat panel, is detected. Antibody binding to H3K27ac in the context of phosphorylation at S28 (H3K27acS28ph) is inhibited to varying degrees in Luminex and ChIP (Figures 1-2).
Histone H3K27ac Antibody, SNAP-ChIP Certified Immunogen: A synthetic peptide corresponding to histone H3 acetylated at lysine 27.

Histone H3K27ac Antibody, SNAP-ChIP Certified Formulation: Protein A affinity-purified antibody (1 mg/mL) in PBS pH 7.4, with 0.05% sodium azide.

Histone H3K27ac Antibody, SNAP-ChIP Certified Storage and Stability: Stable for 1 year at -20°C from date of receipt.

Histone H3K27ac Antibody, SNAP-ChIP Certified Application Notes
Recommended dilutions:
ChIP/ChIP-seq : 2 - 5 μg per 1x10 6 cells
L : 1:4000 dilution
IF : 1:500

Histone H3K27ac Antibody, SNAP-ChIP Certified References:

Grzybowski et al (2015) Mol Cell 58:886

Shah et al (2018) Mol Cell 72:162

View technical datasheet for this product. 13-0045 Datasheet

Applications Key: ChIP-Chromatin IP; E-ELISA; FACS-Flow cytometry; IF-Immunofluorescence; IHC-Immunohistochemistry; ICC-Immunocytochemistry; L-Luminex; 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

Representative SNAP-ChIP-seq results

Representative SNAP-ChIP-seq results: Cumulative histogram plot and heatmap of signal intensity depict H3K27ac ChIP-seq data aligned to annotated transcription start sites (TSS, +/- 3.0 kb; left). Two representative genomic regions depicting H3K27ac peak structure and distribution are shown (right). Data shown are representative of H3K27ac ChIP antibody (EpiCypher Catalog No. 13-0045) 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-AcylStat Spike-in (Catalog No. 19-3001) 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)
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13-0045 Luminex Data

Luminex Data: Histone H3K27ac antibody was assessed using a Luminex® based approach employing dCypher™ Nucleosome K-AcylStat Panel (EpiCypher Catalog No. 16-9003). The panel comprises biotinylated designer nucleosomes (X-Axis) individually coupled to uniquely identifiable Luminex Magplex® beads. Antibody binding to nucleosomes was tested in multiplex (23-plex) at a 1:4000 dilution, and detected with second layer anti-IgG*PE. Data was generated using a Luminex FlexMAP3D®. Data normalized to relevant on-target (H3K27ac; set to 100) is shown.
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13-0045 SNAP-ChIP qPCR Data

SNAP-ChIP-qPCR Data: Histone H3K27ac antibody (3 µg) was tested in a native ChIP experiment using chromatin from K-562 cells (3 µg) with the SNAP-ChIP K-AcylStat Panel (EpiCypher Catalog No. 19-3001) 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)

13-0045 Immunofluorescence

Immunofluorescence Data: IF detection of H3K27ac in HeLa cells stained with H3K27ac antibody at a dilution of 1:500, followed by the addition of an anti-mouse secondary antibody conjugated to a fluorophore (left). The middle pannel shows staining of the nuclei with DAPI. A merge of the two stainings is shown on the right.
(Click image to enlarge)

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