Flow cytometric analysis of HeLa cells, untreated (blue) or treated with UV (100mJ/cm2, 2 hr recovery; green) using Phospho-Chk2 (Thr68) (C13C1) Rabbit mAb (solid lines) or concentration-matched Rabbit (DA1E) mAb IgG XP® Isotype Control #3900 (dashed lines). Anti-rabbit IgG (H+L), F(ab')2 Fragment (Alexa Fluor® 488 Conjugate) was used as a secondary antibody.
Confocal immunofluorescent analysis of HCT 116 (left) and HCT-15 (right) cells using Chk2 (D9C6) XP® Rabbit mAb (green). Actin filaments were labeled with DY-554 phalloidin (red).
Western blot analysis of extracts from control HeLa cells (lane 1) or HeLa cells with a targeted mutation in the gene encoding Chk2 (lane 2) using Chk2 (D9C6) XP® Rabbit mAb (upper) or β-Actin (D6A8) Rabbit mAb #8457 (lower). The change in Chk2 molecular weight in the mutated HeLa cells confirms the specificity of the antibody for Chk2.
Immunohistochemical analysis of paraffin-embedded human breast carcinoma using Phospho-Chk2 (Thr68) (C13C1) Rabbit mAb.
Western blot analysis of extracts from various cell lines using Chk2 (D9C6) XP® Rabbit mAb.
Immunohistochemical analysis of paraffin-embedded human lung carcioma using Chk2 (D9C6) XP® Rabbit mAb.
Immunohistochemical analysis of paraffin-embedded human colon carcinoma, control (left) or λ phosphatase-treated (right), using Phospho-Chk2 (Thr68) (C13C1) Rabbit mAb.
Immunohistochemical analysis of paraffin-embedded human ovarian serous adenocarcinoma using Chk2 (D9C6) XP® Rabbit mAb.
Immunohistochemical analysis of paraffin-embedded human lung carcinoma using Phospho-Chk2 (Thr68) (C13C1) Rabbit mAb.
Immunohistochemical analysis of paraffin-embedded cell pellets, HCT 116 (left) or MDA-MB-231 (right), using Chk2 (D9C6) XP® Rabbit mAb.
Immunohistochemical analysis of paraffin-embedded HT-29 cell pellets, control (left) or UV-treated (right), using Phospho-Chk2 (Thr68) (C13C1) Rabbit mAb.
Immunoprecipitation of Chk2 from 293 cell extracts using Chk2 (D9C6) XP® Rabbit mAb (lane 2). Western blot detection was performed using the same antibody. Lane 1 is 10% input.
Immunoprecipitation of phospho-chk2 from UV-treated HT29 cells using Phospho-Chk2 (Thr68) (C13C1) Rabbit mAb followed by western blot using the same antibody.
Western blot analysis of extracts from HeLa cells, untreated or UV-treated, using Phospho-Chk2 (Thr68) (C13C1) Rabbit mAb.
PhosphoPlus® Duets from Cell Signaling Technology (CST) provide a means to assess protein activation status. Each Duet contains an activation-state and total protein antibody to your target of interest. These antibodies have been selected from CST's product offering based upon superior performance in specified applications.
Chk2 is the mammalian orthologue of the budding yeast Rad53 and fission yeast Cds1 checkpoint kinases (1-3). The amino-terminal domain of Chk2 contains a series of seven serine or threonine residues (Ser19, Thr26, Ser28, Ser33, Ser35, Ser50, and Thr68) each followed by glutamine (SQ or TQ motif). These are known to be preferred sites for phosphorylation by ATM/ATR kinases (4,5). After DNA damage by ionizing radiation (IR), UV irradiation, or hydroxyurea treatment, Thr68 and other sites in this region become phosphorylated by ATM/ATR (5-7). The SQ/TQ cluster domain, therefore, seems to have a regulatory function. Phosphorylation at Thr68 is a prerequisite for the subsequent activation step, which is attributable to autophosphorylation of Chk2 at residues Thr383 and Thr387 in the activation loop of the kinase domain (8).
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