Western blotting analysis using checkpoint kinase 2 antibody (R020839). Total cell lysates (30 μg) from various cell lines were loaded and separated by SDS-PAGE. The blot was incubated with checkpoint kinase 2 antibody (R020839, 1:5,000) and HRP-conjugated goat anti-rabbit secondary antibody (1:20,000) respectively. Image was developed using ECL Substrate Kit.

Western blotting analysis using checkpoint kinase 2 antibody (R020839). Checkpoint kinase 2 expression in wild type (WT) and checkpoint kinase 2 (CHEK2) shRNA knockdown (KD) HeLa cells with 20 μg of total cell lysates. Hsp90 α serves as a loading control. The blot was incubated with checkpoint kinase 2 antibody (R020839, 1:5,000) and HRP-conjugated goat anti-rabbit secondary antibody (1:20,000) respectively. Image was developed using ECL Substrate Kit.

Immunocytochemical staining of Hela cells using Checkpoint kinase 2 antibody (R020839, 1:1,000), Top panel: wild-type (WT); Bottom panal: Checkpoint kinase 2 shRNA knockdown (KD). Nuclei were stained blue with DAPI; Checkpoint kinase 2 was stained magenta with Alexa Fluor® 647. Scale bar, 20 μm.

Validation of Checkpoint kinase 2 knockdown using flow cytometry. Wild-type(WT, Blue) and knockdown(KD, Green) HeLa cells were stained with Checkpoint kinase 2 antibody (R020839, 1:2,000) and analyzed using BD flow cytometer.

Flow cytometric analysis of Checkpoint kinase 2 expression in HAP-1 cells using Checkpoint kinase 2 antibody (R020839, 1:2,000). Green, isotype control; red, Checkpoint kinase 2.

Immunocytochemical staining of HAP1 cells with Checkpoint kinase 2 antibody (R020839, 1:1,000) . Nuclei were stained blue with DAPI;Checkpoint kinase 2 was stained magenta with Alexa Fluor® 647. Images were taken using Leica stellaris 5. Protein abundance based on laser Intensity and smart gain: Medium. Scale bar, 20 μm.

Immunohistochemistry was performed on paraffin-embedded human sigmoid colon carcinoma using checkpoint kinase 2 antibody (R020839, 1:200). Antigen retrieval was done in sodium citrate buffer (pH 6.0). DAB was used for detection, with hematoxylin counterstaining. Images were acquired using a Nikon Ci-L Plus microscope (40× objective). Scale bar: 25 μm.