Histone H3k27ac Antibody, SNAP-Chip® Certified

Histone H3K27ac Antibody, SNAP-ChIP® Certiﬁed Catalog No. 13-0045 Lot No. 21096003-01 Pack Size 50 µg

Type Monoclonal Target Size 15 kDa Reactivity H, M, WR

Host Mouse Format Aff. Pur. IgG Appl. ChIP, ChIP-Seq, L, IF

Product 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 Luminex Data: Histone H3K27ac antibody was assessed using a TM with other lysine acylations in the EpiCypher SNAP-ChIP K- Luminex® based approach employing dCypher Nucleosome K- AcylStat Panel (EpiCypher Catalog No. 16-9003). The panel AcylStat panel is detected. Antibody binding to H3K27ac in the comprises biotinylated designer nucleosomes (x-axis) individually context of phosphorylation at S28 (H3K27acS28ph) is inhibited coupled to uniquely identifiable Luminex MagPlex® beads. to varying degrees in Luminex and ChIP (see figures, right). Antibody binding to nucleosomes was tested in multiplex (23-plex) at a 1:4000 dilution, and detected with second layer anti-IgG*PE. Immunogen: Data was generated using a Luminex FlexMAP3D®. Data A synthetic peptide corresponding to histone H3 acetylated at normalized to relevant on-target (H3K27ac; set to 100) is shown. lysine 27.

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

Storage and Stability: Stable for 1 year at -20°C from date of receipt.

Application Notes: SNAP-ChIP-qPCR. Histone H3K27ac antibody (3 µg) was tested in 6 ChIP: 2 - 5 µg per 1x10 cells a native ChIP experiment using chromatin from K-562 cells (3 µg) with the SNAP-ChIP K-AcylStat Panel (EpiCypher Catalog No. L: 1:4000 19-3001) spiked-in prior to micrococcal nuclease digestion. IF: 1:500 Specificity (left y-axis) was determined by qPCR for the DNA barcodes corresponding to modified nucleosomes in the SNAP- References: ChIP panel (x-axis). Black bar represents antibody efficiency (right Grzybowski et al (2015) Mol Cell 58:886 y-axis; log scale) and indicates percentage of the target Shah et al (2018) Mol Cell 72:162 immunoprecipitated relative to input.

Applications Key: ChIP: Chromatin immunoprecipitation; ChIP-seq: ChIP-sequencing; CUT&RUN: Cleavage Under Targets and Release Using Nuclease; CUT&Tag: Cleavage Under Targets and Tagmentation; E: ELISA; FACS: Flow cytometry; ICC: Immunocytochemistry; IF: Immunoﬂuorescence; IHC: Immunohistochemistry; IP: Immunoprecipitation; L: Luminex; WB: Western Blot. Reactivity Key: B: Bovine; Ce: C. elegans; Ch: Chicken; Dm: Drosophila; Eu: Eukaryote; H: Human; M: Mouse; Ma: Mammal; R: Rat; Sc: S. cerevisiae; Sp: S. pombe; WR: Wide Range (predicted); X: Xenopus; Z: Zebraﬁsh

EpiCypher, Inc • Phone: 855-374-2461 • Fax: 855-420-6111 • www.epicypher.com Immunoﬂuorescence. 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.

EpiCypher, Inc • Phone: 855-374-2461 • Fax: 855-420-6111 • www.epicypher.com