All lanes : Anti-Chk2 (phospho T26) antibody (ab26338) at 1 µg/mlLane 1 : HeLa (Human epithelial carcinoma cell line) Whole Cell Lysate at 10 µgLane 2 : Gamma-Irradiated HeLa Lysate - 4 hour exposure at 20 µgLane 3 : HeLa (Human epithelial carcinoma cell line) Whole Cell Lysate at 10 µg with Human Chk2 (phospho T26) peptide (ab27902) at 1 µg/mlLane 4 : Gamma-Irradiated HeLa Lysate - 4 hour exposure at 20 µg with Human Chk2 (phospho T26) peptide (ab27902) at 1 µg/mlLane 5 : HeLa (Human epithelial carcinoma cell line) Whole Cell Lysate at 10 µg with Human Chk2 peptide (ab37237) at 1 µg/mlLane 6 : Gamma-Irradiated HeLa Lysate - 4 hour exposure at 20 µg with Human Chk2 peptide (ab37237) at 1 µg/mlSecondaryLanes 2 - 6 : IRDye 680 Conjugated Goat Anti-Rabbit IgG (H+L) at 1/15000 dilutionPerformed under reducing conditions.
ab26338 (1/500) staining Chk2 (phospho T26) in HeLa cells (green). Cells were fixed with paraformaldehyde, permeabilised with 0.5% Triton X100/PBS and counterstained with DAPI (blue) in order to highlight the nucleus. The colocalisation of Chk2-pT26 and g-H2AX following bleomycin treatment (induces DNA strand breaks) was confirmed using an anti-gamma H2AX antibody (red). Please refer to abreview for further experimental details.See Abreview