Histone H4K8ac Antibody - SNAP-ChIP® Certified Catalog No. 13-0036 Lot No. 19091001 Pack Size 100 µg Type Monoclonal Host Rabbit Target Size 11.4 kDa Reactivity H, M, WR Format Aff. Pur. IgG Appl. ChIP, ChIP-Seq, WB, ICC, L 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 H4 is one of the four proteins SNAP-ChIP-qPCR: Histone H4K8ac antibody (5 μg) was tested in a native that are present in the nucleosome, the basic repeating subunit ChIP experiment using chromatin from K-562 cells (10 μg) with the SNAP- ChIP K-AcylStat Panel (EpiCypher Catalog No. 19-3001) spiked-in prior to of chromatin, consisting of 147 base pairs of DNA wrapped micrococcal nuclease digestion. Specificity (left y-axis) was determined by around an octamer of core histone proteins (H2A, H2B, H3 and qPCR for the DNA barcodes corresponding to modified nucleosomes in H4). This antibody reacts to H4K8ac, however its binding to the the SNAP-ChIP panel (x-axis). Black bar represents antibody efficiency target is negatively impacted by nearby acetylations (e.g. (right y-axis; log scale) and indicates percentage of the target tetraAc-H4). No cross reactivity with other lysine acylations in immunoprecipitated relative to input. tetraAc-H3 is K4,K9,K14,K18ac; tetraAc-H4 is K5,K8,K12,K16ac; tetraAc- the EpiCypher SNAP-ChIP K-AcylStat panel is detected. H2A is K5,K8,K13,K15ac Immunogen: Synthetic peptide corresponding to histone H4 acetylated at lysine 8. 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. Application Notes: Recommended usage amounts: 6 Luminex Data: Histone H4K8ac antibody was assessed using a Luminex® ChIP 2 - 5 µg per 1x10 cells based approach employing dCypher® Nucleosome K-AcylStat Panel WB 0.5 - 2 µg/mL (EpiCypher Catalog No. 16-9003). The panel comprises biotinylated ICC 0.5 - 2 µg/mL designer nucleosomes (X-Axis) individually coupled to uniquely L 0.25 - 1 µg/mL identifiable Luminex Magplex® beads. Antibody binding to nucleosomes was tested in multiplex (24-plex) at a 1:1000 dilution, and detected with second layer anti-IgG*PE. Data was generated using a Luminex References: FlexMAP3D®. Data normalized to relevant on-target (H4K8ac; set to 100) (1) Shah et al (2018) Mol Cell 72:162 is shown. 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 IP; ChIP-seq: Chromatin IP sequencing E-ELISA; FACS-Flow cytometry; IF-Immunofluorescence; IHC-Immunohistochemistry; ICC-Immunocytochemistry; IP-Immunoprecipitation; WB-Western Blotting; L-Luminex 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 EpiCypher, Inc • Phone: 855-374-2461 • Fax: 855-420-6111 • www.epicypher.com Western Blot Data: Western blot of acid extracts from HeLa cells treated with sodium butyrate (1), acid extracts from untreated HeLa cells (2), and recombinant Histone H4 (3) using EpiCypher Catalog No. 13-0036 at 0.5 µg/mL. Immunocytochemistry Data: Immunocytochemistry of HeLa cells treated with sodium butyrate, using H4K8ac antibody, EpiCypher Catalog No. 13-0036 (red). Actin filaments have been labeled with fluorescein phalloidin (green). Applications Key: ChIP-Chromatin IP; ChIP-seq: Chromatin IP sequencing E-ELISA; FACS-Flow cytometry; IF-Immunofluorescence; IHC-Immunohistochemistry; ICC-Immunocytochemistry; IP-Immunoprecipitation; WB-Western Blotting; L-Luminex 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 EpiCypher, Inc • Phone: 855-374-2461 • Fax: 855-420-6111 • www.epicypher.com.