The protein encoded by this gene is highly similar to the gene product of Schizosaccharomyces pombe rad17, a cell cycle checkpoint gene required for cell cycle arrest and DNA damage repair in response to DNA damage. This protein shares strong similarity with DNA replication factor C (RFC), and can form a complex with RFCs. This protein binds to chromatin prior to DNA damage and is phosphorylated by the checkpoint kinase ATR following damage. This protein recruits the RAD1-RAD9-HUS1 checkpoint protein complex onto chromatin after DNA damage, which may be required for its phosphorylation. The phosphorylation of this protein is required for the DNA-damage-induced cell cycle G2 arrest, and is thought to be a critical early event during checkpoint signaling in DNA-damaged cells. Multiple alternatively spliced transcript variants of this gene, which encode four distinct protein isoforms, have been reported. Two pseudogenes, located on chromosomes 7 and 13, have been identified.

推荐稀释比 WB: 1/1000-1/5000; FC: 1/200-1/2000; ICC/IF: 1/100-1/1000

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Validation of Rad17 knockdown using flow cytometry. Wild-type(WT, Blue) and knockdown(KD, Green) HeLa cells were stained with Phospho-Rad17 (S656) antibody and analyzed using BD flow cytometer.

Flow cytometric analysis of Phospho-Rad17 (S656) expression in HAP-1 cells using Phospho-Rad17 (S656) antibody . Green, isotype control; red, Phospho-Rad17 (S656).

Immunocytochemical staining of HAP-1 cells with Phospho-Rad17 (S656) antibody . Nuclei were stained blue with DAPI; Phospho-Rad17 (S656) 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.

Western blotting analysis using Phospho-Rad17 (S656) antibody . Total cell lysates (30 μg) from various cell lines were loaded and separated by SDS-PAGE. The blot was incubated with Phospho-Rad17 (S656) antibody and HRP-conjugated goat anti-rabbit secondary antibody respectively. Image was developed using FeQ™ ECL Substrate Kit .

Western blotting analysis using phospho-Rad17 (S656) antibody . Phospho-Rad17 (S656) expression in wild-type (WT) and RAD17 shRNA knockdown (KD) HeLa cells with 30 μg of total cell lysates. β-Tubulin serves as a loading control. The blot was incubated with phospho-Rad17 (S656) antibody and HRP-conjugated goat anti-rabbit secondary antibody respectively. Image was developed using NaQ™ ECL Substrate Kit .

Immunocytochemical staining of HeLa cells using Phospho-Rad17 (S656) antibody , Top panel: wild-type (WT); Bottom panal: Phospho-Rad17 (S656) shRNA knockdown (KD). Nuclei were stained blue with DAPI; Phospho-Rad17 (S656) was stained magenta with Alexa Fluor® 647. Scale bar, 20 μm.

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