Histone H3 [Dimethyl Lys18] affinity purified antibody was prepared from whole rabbit serum produced by repeated immunizations with a synthetic dimethylated peptide surrounding Lysine 18 of human Histone H3.2.
0.02 M Potassium Phosphate, 0.15 M Sodium Chloride, pH 7.2
Form:
Liquid (sterile filtered)
Target:
Human
Antibody Type:
Primary Antibody
Application Dilute:
ChIP: 2-5µg/million cells, IHC: 1:50, IF Microscopy: 1:50, WB: 1:500
Application Notes:
Anti-Histone H3 [Dimethyl Lys18] antibody is tested in Western Blot, Chromatin Immunoprecipitation, Dot Blot, and Immunofluorescence. This antibody is useful for Immunocytochemistry. Specific conditions for reactivity should be optimized by the end user.
Chromatin Immunoprecipitation of Rabbit Anti-Histone H3 [Dimethyl Lys18] Antibody. Chromatin from one million formaldehyde cross-linked Hela cells was used with 2ug of Anti-Histone H3 K18me2 and 20ul 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.
Western Blot of Rabbit Anti-Histone H3 [Dimethyl Lys18] Antibody. Lane 1: C. elegans embryo lysate. Load: 30 µg per lane. Primary antibody: Histone H3 [Dimethyl Lys18] at 1:500 for overnight at 4C. Secondary antibody: IRDye800(TM) rabbit secondary antibody at 1:10,000 for 45 min at RT. Block: 5% BLOTTO overnight at 4C. Predicted/Observed size: ~15 kDa. Other band(s): None.
Immunofluorescence of Rabbit Anti-Histone H3 [Dimethyl Lys18] Antibody. Tissue: HeLa cells. Fixation: 0.5% PFA. Antigen retrieval: Not required. Primary antibody: Histone H3 [Dimethyl Lys18] antibody at a 1:500 dilution for 1 h at RT. Secondary antibody: Dylight 488 secondary antibody at 1:10,000 for 45 min at RT. Localization: Histone H3 [Dimethyl Lys18] is nuclear and chromosomal. Staining: Histone H3 [Dimethyl Lys18] is expressed in green, nuclei and alpha-tubulin are counterstained with DAPI (blue) and Dylight 594 (red).
Western Blot of Rabbit Anti-Histone H3 [Dimethyl Lys18] Antibody. Lane 1: NIH-3T3 histone preps. Load: 30 µg per lane. Primary antibody: Histone H3 [Dimethyl Lys18] at 1:500 for overnight at 4C. Secondary antibody: IRDye800(TM) rabbit secondary antibody at 1:10,000 for 45 min at RT. Block: 5% BLOTTO overnight at 4C. Predicted/Observed size: ~15 kDa. Other band(s): None.
Dot Blot of Rabbit Histone H3 [Dimethyl Lys18] Antibody. Lane 1: K18. Lane 2: K18Me2. Lane 3: K18Me3. Lane 4: K18ac. Lane 5: K18Me1. Load: 0.1, 0.3, and 1 picomoles of peptide. Primary antibody: Histone H3 [Dimethyl Lys18] antibody at 1:1000 for 45 min at 4C. Secondary antibody: Dylight(TM)488 rabbit secondary antibody at 1:10,000 for 45 min at RT. Block: 5% BLOTTO overnight at 4C.
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