Histone H3K9ac Antibody, SNAP-ChIP® Certified

SKU: 13-0033
Pack Size: 100 μg
Type:  Monoclonal Host:  Rabbit
Mol Wgt.:  15 kDa Appl.:  ChIP, ChIP-Seq, WB, ICC, E, L
Format:  Aff. Pur. IgG Reactivity: H, M, WR
Histone H3K9ac 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 H3K9ac when present alone as well as in combination with nearby acetylations (tetraAc-H3). No cross reactivity with extended acyl states (H3K9bu, H3K9cr) or other lysine acylations in the EpiCypher SNAP-ChIP K-AcylStat panel, is detected.
Histone H3K9ac Antibody, SNAP-ChIP Certified Certified Immunogen: A synthetic peptide corresponding to histone H3 acetylated at lysine 9.

Histone H3K9ac 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 H3K9ac Antibody, SNAP-ChIP Certified Certified Storage and Stability: Stable for 1 years at -20°C from date of receipt.

Histone H3K9ac Antibody, SNAP-ChIP Certified Certified Application Notes Recommended dilutions:
ChIP: 2 - 5 μg per 1x106 cells WB: 1 - 2 μg/mL ICC: 0.05 - 2 μg/mL E: 0.5 - 2 μg/mL L: 0.25 - 4 μg/mL

Histone H3K9ac Antibody, SNAP-ChIP Certified Certified References:

Grzybowski et al (2015) Mol Cell 58:886

Shah et al (2018) Mol Cell 72:162

View technical datasheet for this product. 13-0033 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

13-0033 SNAP-ChIP Data

Representative SNAP-ChIP-seq results: Cumulative histogram plot and heatmap of signal intensity depict H3K9ac ChIP-seq data aligned to annotated transcription start sites (TSS, +/- 3.0 kb; left). Two representative genomic regions depicting H3K9ac peak structure and distribution are shown (right). Data shown are representative of H3K9ac ChIP antibody (EpiCypher Catalog No. 13-0033) 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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13-0033 SNAP-ChIP Data

SNAP-ChIP qPCR Data: Histone H3K9ac antibody (5 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. tetraAc-H3 is K4,K9,K14,K18ac; tetraAc-H4 is K5,K8,K12,K16ac; tetraAc-H2A is K5,K8,K13,K15ac
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13-0033 Luminex Data

Luminex Data: Histone H3K9ac 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 were tested in multiplex (24-plex) at a 1:1000 dilution, and detected with second layer anti-IgG*PE. Data was generated using a Luminex FlexMAP3D®. Data normalized to relevant on-target (H3K9ac; set to 100) is shown. tetraAc-H3 is K4,K9,K14,K18ac; tetraAc-H4 is K5,K8,K12,K16ac; tetraAc-H2A is K5,K8,K13,K15ac
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13-0033 Western Blot data

Western Blot Data: Western blot of acid extracts from HeLa cells untreated ( - ) or treated with sodium butyrate ( + ), using EpiCypher Catalog No. 13-0033 at 0.25 µg/mL.
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13-0033 ICC

Immunocytochemistry Data: Immunocytochemistry of HeLa cells treated with sodium butyrate, using H3K9ac antibody, EpiCypher Catalog No. 13-0033 (red). Actin filaments have been labeled with fluorescein phalloidin (green).
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13-0033 ELISA Data

ELISA qPCR Data: Sandwich ELISA against H3K9ac using HeLa whole cell lysate, treated or untreated with sodium butyrate, using an anti H3 C-terminal antibody as the capture antibody and EpiCypher Catalog No. 13-0033 as the detection antibody.
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