H3K18me2 | Histone H3 (dimethylated Lys18)
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Chromatin Immunoprecipitation using anti-H3K18me2 antibodies. Chromatin from one million formaldehyde cross-linked Hela cells was used with 2 μg of H3K18me2 antibody and 20 μl of magnetic IgG beads per immunoprecipitation. A no antibody (No Ab) control was also used. Immunoprecipitated DNA was quantified using real-time PCR and normalized to the input chromatin.
Dot Blot using anti-H3K18me2 antibodies. Lane 1: K18 unmodified. Lane 2: K18Me2. Lane 3: K18Me3. Lane 4: K18ac. Lane 5: K18Me1. Load: 0.1, 0.3, and 1 picomoles of peptide. Primary antibody used at a 1:1 000 dilution for 45 min at 4°C. Secondary antibody: Dylight®488 rabbit secondary antibody at 1:10 000 for 45 min at RT.
Immunofluorescence using anti-H3K18me2 antibodies. Tissue: HeLa cells. Fixation: 0.5% PFA. Primary antibody used 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 H3K18me2 is nuclear and chromosomal. Staining: Histone H3K18me2 is expressed in green, nuclei and alpha-tubulin are counterstained with DAPI (blue) and Dylight® 594 (red).
Western Blot using anti-H3K18me2 antibodies. 30 μg of NIH-3T3 histone extract. Primary antibody used at a 1:500 dilution overnight at 4°C. Secondary antibody: IRDye800™ rabbit secondary antibody at 1:10 000 for 45 min at RT.
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