This antibody gave a positive result in ELISA against the immunizing peptide ab26342 (red line). It gave a negative result in ELISA against the non-modified equivalent peptide ab13212 (green line). This indicates that it is specific for the modified peptide.
IHC image of Histone H2B (di methyl K5) 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 ab17351, 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.
ab17351 (1/2000) staining Histone H2B (di methyl K5) in HeLa cells (green). Cells were fixed in paraformaldehyde, permeabilised using 0.5% Triton X100 and counterstained with DAPI in order to highlight the nucleus (red). For further experimental details please refer to Abreview.See Abreview
All lanes : Anti-Histone H2B (di methyl K5) antibody (ab17351) at 1 µg/mlLane 1 : Calf Thymus Histone Preparation Nuclear LysateLane 2 : Calf Thymus Histone Preparation Nuclear Lysate with Human Histone H2B (di methyl K5) peptide (ab26342) at 1 mg/mlLane 3 : Calf Thymus Histone Preparation Nuclear Lysate with Human Histone H2B peptide (ab13212) at 1 mg/mlLysates/proteins at 0.1 µg per lane.SecondaryGoat Anti-Rabbit IgG H&L (HRP) preadsorbed (ab97080) at 1/5000 dilution
ICC/IF image of ab17351 stained HeLa cells. The cells were 100% methanol fixed (5 min), permabilised in 0.1% PBS-Tween (20 min) and incubated with the antibody (ab17351, 5µg/ml) for 1h at room temperature. 1%BSA / 10% normal goat serum / 0.3M glycine was used to 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). This antibody also gave a positive IF result in 4%PFA and 100% methanol fixed Hek293, HepG2 and MCF7 cells. However, this Fast-Track antibody is not yet fully characterised. This image represents inconclusive preliminary data.