All lanes : Anti-Histone H3 (acetyl K23) antibody (ab47813) at 1 µg/mlLane 1 : HeLa Histone Preparation Nuclear Lysate - ButyratedLane 2 : HeLa Histone Preparation Nuclear Lysate - Butyrated with Human Histone H3 peptide (ab17163) at 0.5 µg/mlLane 3 : HeLa Histone Preparation Nuclear Lysate - Butyrated with Histone H3 peptide - acetyl K36 at 0.5 µg/mlLane 4 : HeLa Histone Preparation Nuclear Lysate - Butyrated with Human Histone H3 (acetyl K9) peptide (ab16635) at 0.5 µg/mlLane 5 : HeLa Histone Preparation Nuclear Lysate - Butyrated with Histone H3 peptide - acetyl K14 at 0.5 µg/mlLane 6 : HeLa Histone Preparation Nuclear Lysate - Butyrated with Human Histone H3 (acetyl K18) peptide (ab24003) at 0.5 µg/mlLane 7 : HeLa Histone Preparation Nuclear Lysate - Butyrated with Human Histone H3 (acetyl K27) peptide (ab24404) at 0.5 µg/mlLane 8 : HeLa Histone Preparation Nuclear Lysate - Butyrated with Human Histone H3 (acetyl K23) peptide (ab48359) at 0.5 µg/mlLysates/proteins at 2.5 µg per lan
ICC/IF image of ab47813 stained human HeLa cells. The cells were PFA fixed (10 min), permabilised in TBS-T (20 min) and incubated with the antibody (ab47813, 1µg/ml) for 1h at room temperature. 1%BSA / 10% normal serum / 0.3M glycine was used to quench autofluorescence and block non-specific protein-protein interactions. The secondary antibody (green) was Alexa Fluor® 488 goat anti-rabbit IgG (H+L) used at a 1/1000 dilution for 1h. Alexa Fluor® 594 WGA was used to label plasma membranes (red). DAPI was used to stain the cell nuclei (blue).
IHC image of hISTONE h3 (acetyl K23) staining in human breast carcinoma FFPE section, performed on a BondTM system using the standard protocol F. The section was pre-treated using heat mediated antigen retrieval with sodium citrate buffer (pH6, epitope retrieval solution 1) for 20 mins. The section was then incubated with ab47813, 1µg/ml, for 15 mins at room temperature and detected using an HRP conjugated compact polymer system. DAB was used as the chromogen. The section was then counterstained with haematoxylin and mounted with DPX.