Histone H3 K79Me3 antibody

• Catalog Number: orb420422
• Category: Antibodies
• Description: Histone H3 K79Me3 antibody
• Species/Host: Rabbit
• Clonality: Polyclonal
• Tested applications: ChIP, DOT, IF, IHC, Multiplex Assay, WB
• Reactivity: C. elegans, Human
• Isotype: IgG
• Immunogen: Histone H3 [Trimethyl Lys79] affinity purified antibody was prepared from whole rabbit serum produced by repeated immunizations with a synthetic trimethylated peptide surrounding Lysine 79 of human Histone H3.
• Concentration: 0.6 mg/ml
• Dilution range: ChIP: 2-5µg/million cells, IHC: 1:100-1:1000, IF: 1:100-1:1000, WB: 1µg/mL
• Form/Appearance: Liquid (sterile filtered)
• Purity: Anti-Histone H3 [Trimethyl Lys79] was affinity purified from monospecific antiserum by immunoaffinity chromatography. This antibody reacts with human Histone H3. 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: 0.01% (w/v) Sodium Azide
• Alternative names: rabbit anti-Histone H3 trimethyl Lys79 antibody, H
• Note: For research use only
• Application notes: Anti-Histone H3 [Trimethyl Lys79] antibody is tested in Western Blot, Dot Blot, Chromatin Immunoprecipitation, and Immunofluorescence. This antibody is useful for 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.

Chromatin Immunoprecipitation of Rabbit Anti-Histone H3 [Trimethyl Lys79] Antibody. Chromatin from one million formaldehyde cross-linked HeLa cells was used with 2 ug of Anti-Histone H3 K27me3 and 20 ul of magnetic IgG beads per immunoprecipitation. A no antibody (No Ab) control was also used. Immunoprecipitated DNA was quantified using quantitative real-time PCR and SYBR green dye, then normalized to the non-precipitated input chromatin, which is equal to one.

Dot Blot of Rabbit Histone H3 [Trimethyl Lys79] Antibody. Lane 1: K79 unmodified. Lane 2: K79me1. Lane 3: K79me2. Lane 4: K79me3. Load: 1, 10, and 100 picomoles of peptide. Primary antibody: Histone H3 [Trimethyl Lys79] antibody at 1:1000 for 45 min at 4°C. Secondary antibody: Dylight™488 rabbit secondary antibody at 1:10000 for 45 min at RT. Block: 5% BLOTTO overnight at 4°C.

Immunofluorescence of Rabbit Anti-Histone H3 [Trimethyl Lys79] Antibody. Tissue: HeLa cells during prophase. Fixation: 0.5% PFA. Antigen retrieval: Not required. Primary antibody: Histone H3 [Trimethyl Lys79] antibody at a 1:100 dilution for 1 h at RT. Secondary antibody: Dylight 488 secondary antibody at 1:10000 for 45 min at RT. Localization: Histone H3 [Trimethyl Lys79] is nuclear and chromosomal. Staining: Histone H3 [Trimethyl Lys79] is expressed in green, nuclei and alpha-tubulin are counterstained with DAPI (blue) and Dylight 550 (red).

Western Blot of Rabbit Anti-Histone H3 [Trimethyl Lys79] Antibody. Lane 1: HeLa Histone prep. Lane 2: 3T3 Histone prep. Lane 3: C. elegans embryo lysate. Load: 30 µg per lane. Primary antibody: Histone H3 [Trimethyl Lys79] at 1 µg/ml 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.