Histone H3 [p Thr6, Monomethyl Lys9] affinity purified antibody was prepared from whole rabbit serum produced by repeated immunizations with a synthetic monomethylated/phosphorylated peptide surrounding Lysine 9/Threonine 6 of human Histone H3.2.
Preservative: 0.01% (w/v) Sodium Azide. Stabilizer: 30% Glycerol, Buffer: 0.02 M Potassium Phosphate, 0.15 M Sodium Chloride, pH 7.2
Reinheit:
Anti-Histone H3 [p Thr6, Monomethyl Lys9] 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.
Application Notes: Anti-Histone H3 [p Thr6, Monomethyl Lys9] antibody is tested for Western Blot, Immunocytochemistry, Immunofluorescence, Chromatin Immunoprecipitation, and Dot Blot. 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
Chromatin Immunoprecipitation of Histone H3 [p Thr6, Monomethyl Lys9] Antibody. Chromatin from one million formaldehyde cross-linked Hela cells was used with 2 ug of Anti-Histone H3 K9me2pT6 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 [Monomethyl Lys9, p Thr6] Antibody. Lane 1: Histone H3 1-13. Lane 2: T6p. Lane 3: T6pK9Me1. Lane 4: T6pK9Me2. Lane 5: T6pK9Me3. Lane 6: Histone H3 2-17. Lane 7: K9Me1. Lane 8: K9Me2. Lane 9: K9Me3. Lane 10: K9Ac. Load: 1, 10, and 100 picomoles of peptide. Primary antibody: Histone H3 [Monomethyl Lys9, p Thr6] antibody at 1:1000 for 45 min at 4C. Secondary antibody: Dylight(TM)488 rabbit secondary antibody at 1:10000 for 45 min at RT. Block: 5% BLOTTO overnight at 4C.
Immunofluorescence of Rabbit Anti-Histone H3 [p Thr6, Monomethyl Lys9] Antibody. Tissue: HeLa cells. Fixation: 0.5% PFA. Antigen retrieval: Not required. Primary antibody: Histone H3 [p Thr6, Monomethyl Lys9] 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 H3 [p Thr6, Monomethyl Lys9] is nuclear and chromosomal. Staining: Histone H3 [p Thr6, Monomethyl Lys9] is expressed in green and the nuclei are counterstained with DAPI (blue).
Western Blot of Rabbit Anti-Histone H3 [Monomethyl Lys9, p Thr6] Antibody. Lane 1: C. elegans embryo lysate. Load: 30 µg per lane. Primary antibody: Histone H3 [Monomethyl Lys9, p Thr6] at 1:1000 for overnight at 4C. Secondary antibody: IRDye800(TM) rabbit secondary antibody at 1:10000 for 45 min at RT. Block: 5% BLOTTO overnight at 4C. Predicted/Observed size: ~15 kDa. Other band(s): None.
Western Blot of Rabbit Anti-Histone H3 [Monomethyl Lys9, p Thr6] Antibody. Lane 1: HeLa Histone prep lysate. Load: 30 µg per lane. Primary antibody: Histone H3 [Monomethyl Lys9, p Thr6] at 1:1000 for overnight at 4C. Secondary antibody: IRDye800(TM) rabbit secondary antibody at 1:10000 for 45 min at RT. Block: 5% BLOTTO overnight at 4C. Predicted/Observed size: ~15 kDa. Other band(s): None.
Western Blot of Rabbit Anti-Histone H3 [Monomethyl Lys9, p Thr6] Antibody. Lane 1: NIH-3T3 Histone prep lysate. Load: 30 µg per lane. Primary antibody: Histone H3 [Monomethyl Lys9, p Thr6] at 1:1000 for overnight at 4C. Secondary antibody: IRDye800(TM) rabbit secondary antibody at 1:10000 for 45 min at RT. Block: 5% BLOTTO overnight at 4C. Predicted/Observed size: ~15 kDa. Other band(s): None.
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