Histone H3 R2me2s/K4me2 antibody

• Catalog Number: orb420412
• Category: Antibodies
• Description: Histone H3 R2me2s/K4me2 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 [Sym-dimethyl Arg2, Dimethyl Lys4] affinity purified antibody was prepared from whole rabbit serum produced by repeated immunizations with synthetic dimethylated/dimethylated peptides surrounding Arginine 2 / Lysine 4 of human Histone H3.2.
• Concentration: 0.91 mg/ml
• Dilution range: ChIP: 2-5µg/million cells, IHC: 1:100, IF: 1:100, WB: 1:500
• Form/Appearance: Liquid (sterile filtered)
• Purity: Anti-Histone H3 [Sym-dimethyl Arg2, Dimethyl Lys4] 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: 0.01% (w/v) Sodium Azide
• Alternative names: rabbit anti-Histone H3 Sym-dimethyl Arg2 dimethyl
• Note: For research use only
• Application notes: Anti-Histone H3 [Sym-dimethyl Arg2, Dimethyl Lys4] antibody is tested for Western Blot, Chromatin Immunoprecipitation, Dot Blot, and Immunofluorescence. 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 [Sym-dimethyl Arg2, Dimethyl Lys4] Antibody. Chromatin from one million formaldehyde cross-linked Hela cells was used with 2 ug of Anti-Histone H3 R2me2s-K4me2 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 [Sym-dimethyl Arg2, Dimethyl Lys4] Antibody. Lane 1: R2Me2. Lane 2: R2Me2/K4Me2. Lane 3: K4Me2. Load: 0.1, 1, 0.5, 5, and 10 picomoles of peptide. Primary antibody: Histone H3 [Sym-dimethyl Arg2, Dimethyl Lys4] 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. Final Antibody reactivity to R2Me2/K4Me2 seen in the right blot.

Immunofluorescence of Rabbit Anti-Histone H3 [Sym-dimethyl Arg2, Dimethyl Lys4] Antibody. Tissue: HeLa cells. Fixation: 0.5% PFA. Antigen retrieval: Not required. Primary antibody: Histone H3 [Sym-dimethyl Arg2, Dimethyl Lys4] 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 [Sym-dimethyl Arg2, Dimethyl Lys4] is nuclear and chromosomal. Staining: Histone H3 [Sym-dimethyl Arg2, Dimethyl Lys4] is expressed in green, nuclei and alpha-tubulin are counterstained with DAPI (blue) and Dylight 594 (red).

Western Blot of Rabbit Anti-Histone H3 [Sym-dimethyl Arg2, Dimethyl Lys4] Antibody. Lane 1: C. elegans embryo cell lysate. Load: 30 µg per lane. Primary antibody: Histone H3 [Sym-dimethyl Arg2, Dimethyl Lys4] 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 [Sym-dimethyl Arg2, Dimethyl Lys4] Antibody. Lane 1: NIH-3T3 histone preps. Load: 30 µg per lane. Primary antibody: Histone H3 [Sym-dimethyl Arg2, Dimethyl Lys4] 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.