Histone H3K9ac Antibody, SNAP-ChIP® Certified Catalog No. 13-0033 Lot No. 21125003-01 Pack Size 100 µg Type Monoclonal Host Rabbit Target Size 15 kDa Reactivity H, M, WR Format Aff. Pur. IgG Appl. ChIP, ChIP-Seq, WB, ICC, E, L Product Description: This antibody meets EpiCypher’s “SNAP-ChIP® Certified” criteria for SNAP-ChIP-qPCR: Histone H3K9ac antibody (3 μg) was tested in specificity and efficient target enrichment in a ChIP experiment (<20% a native ChIP experiment using chromatin from K-562 cells (3 μg) cross-reactivity across the panel, >5% recovery of target input). with the SNAP-ChIP K-AcylStat Panel (EpiCypher Catalog No. Histone H3 is one of the four proteins that are present in the 19-3001) spiked-in prior to micrococcal nuclease digestion. nucleosome, the basic repeating subunit of chromatin, consisting of Specificity (left y-axis) was determined by qPCR for the DNA 147 base pairs of DNA wrapped around an octamer of core histone barcodes corresponding to modified nucleosomes in the SNAP- proteins (H2A, H2B, H3 and H4). This antibody reacts to H3K9ac ChIP K-AcylStat panel (x-axis). Black bar represents antibody when present alone as well as in combination with nearby acetylations efficiency (right y-axis; log scale) and indicates percentage of the (tetraAc-H3). No cross reactivity with extended acyl states (H3K9bu, target immunoprecipitated relative to input. H3K9cr) or other lysine acylations in the EpiCypher SNAP-ChIP K- tetraAc-H3 is K4,K9,K14,K18ac; tetraAc-H4 is K5,K8,K12,K16ac; AcylStat panel is detected. tetraAc-H2A is K5,K8,K13,K15ac Immunogen: Synthetic peptide corresponding to histone H3 acetylated lysine 9. Formulation: Protein A affinity-purified antibody (1 mg/mL) in PBS, with 0.09% sodium azide, 1% BSA, and 50% glycerol. Storage and Stability: Stable for 1 year at -20°C from date of receipt. Luminex Data: Histone H3K9ac antibody was assessed using a Luminex® based approach employing dCypherTM Nucleosome K- Application Notes: AcylStat Panel (EpiCypher Catalog No. 16-9003). The panel Recommended Dilutions: comprises biotinylated designer nucleosomes (X-Axis) individually ChIP: 2 - 5 µg per 1x106 cells WB: 0.25 - 1 µg/mL coupled to uniquely identifiable Luminex MagPlex® beads. Antibody ICC: 0.5 - 2 µg/mL E: 0.5 - 2 µg/mL binding to nucleosomes were tested in multiplex (24-plex) at a L: 0.25 - 4 µg/mL 1:250 dilution, and detected with second layer anti-IgG*PE. Data was generated using a Luminex FLEXMAP 3D®. Data normalized to References: relevant on-target (H3K9ac; set to 100) is shown. (1) Shah et al (2018) Mol Cell 72:162 tetraAc-H3 is K4,K9,K14,K18ac; tetraAc-H4 is K5,K8,K12,K16ac; tetraAc-H2A is K5,K8,K13,K15ac Applications Key: ChIP: Chromatin immunoprecipitation; 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 This product is for in vitro research use only and is not intended for use in humans or animals. EpiCypher, Inc • Phone: 855-374-2461 • Fax: 855-420-6111 • www.epicypher.com Western Blot Data: Western blot of acid extracts from HeLa cells untreated ( - ) or treated with sodium butyrate ( + ), using H3K9ac antibody (EpiCypher Catalog No. 13-0033) at 0.25 µg/mL. 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). ELISA 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 H3K9ac antibody (EpiCypher Catalog No. 13-0033) as the detection antibody. Applications Key: ChIP: Chromatin immunoprecipitation; 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 This product is for in vitro research use only and is not intended for use in humans or animals. EpiCypher, Inc • Phone: 855-374-2461 • Fax: 855-420-6111 • www.epicypher.com.