Histone H3 K9ac/K14ac antibody

• Catalog Number: orb420400
• Category: Antibodies
• Description: Histone H3 K9ac/K14ac antibody
• Species/Host: Rabbit
• Clonality: Polyclonal
• Tested applications: ChIP, DOT, IF, IHC, Multiplex Assay, WB
• Reactivity: C. elegans, Human, Mouse
• Isotype: IgG
• Immunogen: Histone H3 [ac Lys9, ac Lys14] affinity purified antibody was prepared from whole rabbit serum produced by repeated immunizations with a synthetic acetylated peptide surrounding Lysines 9 and 14 of human Histone H3.2.
• Concentration: 0.63mg/ml
• Dilution range: ChIP: 2-5µg/million cells, IHC: 1:200, IF: 1:200, WB: 1:500
• Form/Appearance: Liquid (sterile filtered)
• Purity: Anti-Histone H3 [ac Lys9, ac Lys14] was affinity purified from monospecific antiserum by immunoaffinity chromatography. This antibody reacts with human Histone H3.2. A BLAST analysis was used to suggest cross-reactivity with Human, mouse, and C. elegans. Predicted to react with many species including rat, chicken, Xenopus, Drosophila, and plant based on 100% sequence homology. Cross-reactivity with Histone H3 from other sources has not been determined.
• Conjugation: Unconjugated
• UniProt ID: Q71DI3
• NCBI: NP_001005464
• Storage: Store vial at -20° C prior to opening. Aliquot contents and freeze at -20° C or below for extended storage. Avoid cycles of freezing and thawing. Centrifuge product if not completely clear after standing at room temperature. This product is stable for several weeks at 4° C as an undiluted liquid. Dilute only prior to immediate use.
• Buffer/Preservatives: 0.05% (w/v) Sodium Azide
• Alternative names: rabbit anti-Histone H3 Ac Lys9 / Ac Lys14 antibody
• Note: For research use only
• Application notes: Anti-Histone H3 [ac Lys9, ac Lys14] antibody is tested for Western Blot, Dot Blot, and Immunofluorescence. This antibody is useful for Chromatin Immunoprecipitation and Immunocytochemistry. Specific conditions for reactivity should be optimized by the end user. Expect a band approximately ~15.4 kDa corresponding to Histone H3 protein by Western Blotting in HeLa histone prep lysate or the appropriate cell lysate or extract. Epi-Plus antibody production in collaboration with Novus Biologicals.
• Expiration Date: 12 months from date of receipt.

Dot Blot of Rabbit Histone H3 [K9ac, K14ac] Antibody. Lane 1: K9-K14. Lane 2: K9-KAc-K14-Kac. Lane 3: K9-K14-Kac. Lane 4: K9-Ac-K14. Load: 0.3, 1, and 3 µg of peptide. Primary antibody: Histone H3 [ac Lys9, ac Lys14] antibody for 45 min at 4°C. Secondary antibody: HRP rabbit secondary antibody at 1:10000 for 45 min at RT. Block: 5% BLOTTO A overnight at 4°C.

Immunofluorescence of Rabbit Anti-Histone H3 [ac Lys9, ac Lys14] Antibody. Tissue: HeLa cells. Fixation: 0.5% PFA. Antigen retrieval: Not required. Primary antibody: Histone H3 [ac Lys9, ac Lys14] antibody at a 1:200 dilution for 1 h at RT. Secondary antibody: FITC secondary antibody at 1:10000 for 45 min at RT. Localization: Histone H3 [ac Lys9, ac Lys14] is nuclear and chromosomal. Staining: Histone H3 [ac Lys9, ac Lys14] is expressed in green, nuclei and actin are counterstained with Dapi (blue) and Phalloidin (red).

ImmunoFluorescence of Rabbit Anti-Histone H3 K9ac/K14ac Antibody. Cells: HeLa Cells. Fixation: 4% PFA. Permeabilization: 0.3% Triton X-100. Primary Antibody: Anti-Histone H3 [ac Lys9, ac Lys14] at 5 µg/mL overnight at 2-8°C. Secondary Antibody: Goat Anti-Rabbit IgG DyLight™488 at 5 µg/mL for 1 hr at RT. Nuclear Counterstain: DAPI. Actin Filament Stain: Texas Red - X Phallodin. Staining: (A) DAPI. (B) K9 AC/K14 AC + DyLight™488. (C) Actin-X Phallodin. (D) Merge A+ B. (E) Merge A+ B+ C. (F) Secondary Only Merge. Expected Location: Nuclear and Chromosomal.

Western Blot of Rabbit Anti-Histone H3 [ac Lys9, ac Lys14] Antibody. Lane 1: C. elegans embryo lysate. Load: 30 µg per lane. Primary antibody: Histone H3 [ac Lys9, ac Lys14] at 1:500 for overnight at 4°C. Secondary antibody: IRDye800™ rabbit secondary antibody at 1:10000 for 45 min at RT. Block: 5% BLOTTO overnight at 4°C. Predicted/Observed size: ~15 kDa. Other band(s): None.

Western Blot of Rabbit Anti-Histone H3 [ac Lys9, ac Lys14] Antibody. Lane 1: NIH-3T3 histone preps. Load: 30 µg per lane. Primary antibody: Histone H3 [ac Lys9, ac Lys14] at 1:500 for overnight at 4°C. Secondary antibody: IRDye800™ rabbit secondary antibody at 1:10000 for 45 min at RT. Block: 5% BLOTTO overnight at 4°C. Predicted/Observed size: ~15 kDa. Other band(s): None.