All lanes : Anti-Chk2 (phospho S28) antibody (ab26340) at 1 µg/mlLane 1 : HeLa Gamma Irradiated Whole Cell Lysate Lane 2 : HeLa Gamma Irradiated Whole Cell Lysate with Human Chk2 (phospho S28) peptide (ab28701) at 1 µg/mlLane 3 : HeLa Gamma Irradiated Whole Cell Lysate with Human Chk2 peptide (ab28702) at 1 µg/mlLysates/proteins at 20 µg per lane.SecondaryIRDye 680 Conjugated Goat Anti-Rabbit IgG (H+L) at 1/10000 dilutionPerformed under reducing conditions.
ICC/IF image of ab26340 stained human HeLa cells. The cells were 4% PFA fixed (10 min), permabilised in TBS-T (20 min) and incubated with the antibody (ab26340, 1µg/ml) for 1h at room temperature. 1%BSA / 10% normal goat 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).
ab26340 (1/500) staining Chk2 (phospho S28) in HeLa cells. Cells were pre-treated with Bleomycin for 2 hours, paraformaldehyde fixed and permeabilised with 0.5% Triton X100/PBS. Colocalisation with sites of double-strand DNA repair was confirmed through the use of an anti-gamma H2AX antibody (ab18311). Please refer to abreview for further experimental details.See Abreview