In response to DNA damage and replication blocks, cell cycle progression is halted through the control of critical cell cycle regulators. The protein encoded by this gene is a cell cycle checkpoint regulator and putative tumor suppressor. It contains a forkhead-associated protein interaction domain essential for activation in response to DNA damage and is rapidly phosphorylated in response to replication blocks and DNA damage. When activated, the encoded protein is known to inhibit CDC25C phosphatase, preventing entry into mitosis, and has been shown to stabilize the tumor suppressor protein p53, leading to cell cycle arrest in G1. In addition, this protein interacts with and phosphorylates BRCA1, allowing BRCA1 to restore survival after DNA damage. Mutations in this gene have been linked with Li-Fraumeni syndrome, a highly penetrant familial cancer phenotype usually associated with inherited mutations in TP53. Also, mutations in this gene are thought to confer a predisposition to sarcomas, breast cancer, and brain tumors. This nuclear protein is a member of the CDS1 subfamily of serine/threonine protein kinases. Several transcript variants encoding different isoforms have been found for this gene.

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Western blotting analysis using Checkpoint kinase 2 antibody . 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 and HRP-conjugated goat anti-rabbit secondary antibody respectively. Image was developed using FeQ™ ECL Substrate Kit .

Western blotting analysis using Checkpoint kinase 2 antibody . Checkpoint kinase 2 expression in wild type (WT) and Checkpoint kinase 2 shRNA knockdown (KD) HeLa cells with 30 μg of total cell lysates. Hsp90 α serves as a loading control. The blot was incubated with Checkpoint kinase 2 antibody and HRP-conjugated goat anti-rabbit secondary antibody respectively. Image was developed using FeQ™ ECL Substrate Kit .

Flow cytometric analysis of Checkpoint kinase 2 expression in HepG2 cells using Checkpoint kinase 2 antibody . Green, isotype control; red, Checkpoint kinase 2.

Immunocytochemical staining of HT-1080 cells with Checkpoint kinase 2 antibody . 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.

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 and analyzed using BD flow cytometer.

Immunocytochemical staining of Hela cells using Checkpoint kinase 2 antibody , 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.

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