HIST4H4 Antibody

SKU: orb420429

Description

Histone H4 K20me2 antibody

Images & Validation

• Tested Applications: ChIP, DOT, IF, IHC, WB
• Dilution Range: ChIP: 2-5µg/million cells, IHC: 1:50, IF: 1:50, WB: 1:2000
• Reactivity: C. elegans, Human
• Application Notes:
  Anti-Histone H4 [Dimethyl Lys20] antibody is tested for Western Blot, Dot Blot, Chromatin Immunoprecipitation, Immunofluorescence, and Immunocytochemistry. Specific conditions for reactivity should be optimized by the end user. Expect a band approximately ~13 kDa corresponding to Histone H4 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.

Key Properties

• Antibody Type: Primary Antibody
• Host: Rabbit
• Clonality: Polyclonal
• Isotype: IgG
• Immunogen: Histone H4 [Dimethyl Lys20] affinity purified antibody was prepared from whole rabbit serum produced by repeated immunizations with a synthetic dimethylated peptide surrounding Lysine 20 of human Histone H4.
• Target: HIST4H4
• Purity: Anti-Histone H4 [Dimethyl Lys20] was affinity purified from monospecific antiserum by immunoaffinity chromatography. This antibody reacts with human Histone H4. A BLAST analysis was used to suggest cross-reactivity with Human, mouse, and C. elegans. Predicted to react with most mammal species. Cross-reactivity with Histone H4 from other sources has not been determined.
• Conjugation: Unconjugated

Storage & Handling

• 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.
• Form/Appearance: Liquid (sterile filtered)
• Buffer/Preservatives: Preservative: 0.01% (w/v) Sodium Azide. Stabilizer: 30% Glycerol; Buffer: 0.02 M Potassium Phosphate, 0.15 M Sodium Chloride, pH 7.2
• Concentration: 0.53 mg/ml
• Expiration Date: 12 months from date of receipt.
• Dry Ice Shipping: Please note: This product requires shipment on dry ice. A dry ice surcharge will apply.
• Disclaimer: For research use only

Alternative Names

rabbit anti-Histone H4 dimethyl Lys20 antibody, H4K20me2, HIST2H4B, HIST1H4H, HIST1H4I, HIST1H4J, HIST1H4K, HIST1H4L, HIST2H4, HIST2H4A, HIST1H4A, HIST1H4B, HIST1H4C, HIST1H4D, HIST1H4E, HIST1H4F, histone cluster 4, H4, histone 4, H4

Chromatin Immunoprecipitation of Rabbit Anti-Histone H4 [Dimethyl Lys20] Antibody. Chromatin from one million formaldehyde cross-linked Hela cells was used with 2 ug of Anti-Histone H4 K20me2 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 H4 [Dimethyl Lys20] 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, 3, 10, and 30 picomoles of peptide. Primary antibody: Histone H4 [Dimethyl Lys20] antibody at 1:2000 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 H4 [Dimethyl Lys20] Antibody. Tissue: HeLa cells. Fixation: 0.5% PFA. Antigen retrieval: Not required. Primary antibody: Histone H4 [Dimethyl Lys20] antibody at a 1:50 dilution for 1 h at RT. Secondary antibody: FITC secondary antibody at 1:10000 for 45 min at RT. Localization: Histone H4 [Dimethyl Lys20] is nuclear and chromosomal. Staining: Histone H4 [Dimethyl Lys20] is expressed in green, nuclei are counterstained with DAPI (blue).

Western Blot of Rabbit Anti-Histone H4 [Dimethyl Lys20] Antibody. Lane 1: C. elegans embryo lysate. Load: 30 µg per lane. Primary antibody: Histone H4 [Dimethyl Lys20] at 1:2000 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: ~13 kDa. Other band(s): None.

Western Blot of Rabbit Anti-Histone H4 [Dimethyl Lys20] Antibody. Lane 1: HeLa histone preps. Load: 30 µg per lane. Primary antibody: Histone H4 [Dimethyl Lys20] at 1:2000 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: ~13 kDa. Other band(s): None.

Quick Database Links

Gene Symbol

HIST4H4

UniProt

P62805

RefSeq (Protein):NP_001029249