Histone H4 K20me1 antibody

• Catalog Number: orb420428
• Category: Antibodies
• Description: Histone H4 K20me1 antibody
• Species/Host: Rabbit
• Clonality: Polyclonal
• Tested applications: ChIP, DOT, IF, IHC, Multiplex Assay, WB
• Reactivity: C. elegans, Human, Mouse
• Isotype: IgG
• Immunogen: Histone H4 [Monomethyl Lys20] affinity purified antibody was prepared from whole rabbit serum produced by repeated immunizations with a synthetic monomethylated peptide surrounding Lysine 20 of human Histone H4.
• Concentration: 1.12 mg/ml
• Dilution range: ChIP: 2-5µg/million cells, IHC: 1:50, IF: 1:50, WB: 1:500
• Form/Appearance: Liquid (sterile filtered)
• Purity: Anti-Histone H4 [Monomethyl 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
• UniProt ID: P62805
• NCBI: NP_001029249
• 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 H4 monomethyl Lys20 antibody,
• Note: For research use only
• Application notes: Anti-Histone H4 [Monomethyl 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.
• Expiration Date: 12 months from date of receipt.

Chromatin Immunoprecipitation Rabbit Anti-Histone H4 [Monomethyl Lys20] Antibody. Chromatin from one million formaldehyde cross-linked Hela cells was used with 2 ug of Anti-Histone H4K20me1 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 [Monomethyl Lys20] Antibody. Antigen: Lane 1: unmodified. Lane 2: K20me1. Lane 3: K20me2. Lane 4: K20me3. Load: 0.6, 1, 3, 6, and 10 picomoles of peptide. Primary antibody: Histone H4 [Monomethyl Lys20] antibody at 2 µg/ml 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 [Monomethyl Lys20] Antibody. Tissue: HeLa cells. Fixation: 0.5% PFA. Antigen retrieval: Not required. Primary antibody: Histone H4 [Monomethyl Lys20] antibody at a 1:500 dilution for 1 h at RT. Secondary antibody: FITC secondary antibody at 1:10000 for 45 min at RT. Localization: Histone H4 [Monomethyl Lys20] is nuclear and chromosomal. Staining: Histone H4 [Monomethyl Lys20] is expressed in green, nuclei and alpha-tubulin are counterstained with DAPI (blue) and Dylight 550 (red).

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

Western Blot of Rabbit Anti-Histone H4 [Monomethyl Lys20] Antibody. Lane 1: NIH-3T3 histone preps. Load: 30 µg per lane. Primary antibody: Histone H4 [Monomethyl Lys20] 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: ~13 kDa. Other band(s): None.