Cell Signaling Technology

Product Pathways - DNA Damage

Phospho-Chk1 (Ser345) (133D3) Rabbit mAb (PE Conjugate) #12268

Applications Dilution Species-Reactivity Sensitivity MW (kDa) Isotype
F 1:50 Human,Mouse,Rat,Monkey, Endogenous Rabbit IgG

Species cross-reactivity is determined by western blot.

Applications Key: F=Flow Cytometry,

Specificity / Sensitivity

Phospho-Chk1 (Ser345) (133D3) Rabbit mAb (PE Conjugate) detects endogenous levels of Chk1 protein only when phosphorylated at Ser345.Phospho-Chk1 (Ser345) (133D3)Rabbit mAb(PE偶联)可以检测到345位丝氨酸磷酸化的内源性Chk1蛋白。

Source / Purification

Monoclonal antibody is produced by immunizing animals with a synthetic phosphopeptide corresponding to residues surrounding Ser345 of human Chk1 protein.单克隆抗体由合成肽段免疫动物产生,该肽段与人Chk1的345位丝氨酸邻近残基序列一致。


This Cell Signaling Technology antibody is conjugated to phycoerythrin (PE) and tested in-house for direct flow cytometry analysis in human cells. The antibody is expected to exhibit the same species cross-reactivity as the unconjugated Phospho-Chk1 (Ser345) (133D3) Rabbit mAb #2348. Cell Signaling Technology公司抗体偶联藻红蛋白(PE)并经人类细胞直接流式细胞仪免疫荧光分析内部测试。抗体预期和未偶联的Phospho-Chk1 (Ser345) (133D3) 兔 mAb #2348显示同样的物种交叉反应。

Flow Cytometry

Flow Cytometry

Flow cytometric analysis of HeLa cells, untreated (blue) and UV-treated (green), using Phospho-Chk1 (Ser345) (133D3) Rabbit mAb (PE Conjugate).使用Phospho-Chk1 (Ser345) (133D3)Rabbit mAb(PE偶联)对未处理(蓝色)和UV-处理(绿色)的HeLa细胞进行流式细胞分析。


Chk1 kinase acts downstream of ATM/ATR kinase and plays an important role in DNA damage checkpoint control, embryonic development, and tumor suppression (1). Activation of Chk1 involves phosphorylation at Ser317 and Ser345 and occurs in response to blocked DNA replication and certain forms of genotoxic stress (2). While phosphorylation at Ser345 serves to localize Chk1 to the nucleus following checkpoint activation (3), phosphorylation at Ser317 along with site-specific phosphorylation of PTEN allows for re-entry into the cell cycle following stalled DNA replication (4). Chk1 exerts its checkpoint mechanism on the cell cycle, in part, by regulating the cdc25 family of phosphatases. Chk1 phosphorylation of cdc25A targets it for proteolysis and inhibits its activity through 14-3-3 binding (5). Activated Chk1 can inactivate cdc25C via phosphorylation at Ser216, blocking the activation of cdc2 and transition into mitosis (6). Centrosomal Chk1 has been shown to phosphorylate cdc25B and inhibit its activation of CDK1-cyclin B1, thereby abrogating mitotic spindle formation and chromatin condensation (7). Furthermore, Chk1 plays a role in spindle checkpoint function through regulation of aurora B and BubR1 (8). Research studies have implicated Chk1 as a drug target for cancer therapy as its inhibition leads to cell death in many cancer cell lines (9).Chk1激酶是ATM/ATR激酶的下游,在DNA损伤检验点控制,胚胎发育和癌症抑制过程中发挥了重要作用(1)。Chk1的激活包括317位和345位丝氨酸的磷酸化,这一过程是对DNA复制抑制和某些基因毒性压力的应激(2)。345位丝氨酸磷酸化可以帮助Chk1在检验点激活后定位到细胞核(3),317位磷酸化和PTEN某些特定位点磷酸化可以是细胞在停滞的DNA复制后重新进入细胞周期(4)。Chk1通过部分调控磷酸酯酶cdc25家族蛋白,发挥它对细胞周期检验点的调控作用。Chk1磷酸化cdc25A促进它的蛋白酶解并通过14-3-3结合抑制它的活性(5)。激活的Chk1可以通过磷酸化cdc25C使其失活,阻断cdc2的活性使其无法进入细胞周期(6)。中心体Chk1已经证明可以磷酸化cdc25B,也能抑制cdc25B对CDK1-cyclin B1的激活作用,从而废除有丝分裂纺锤体的形成和染色质缩合(7)。此外,Chk1通过调节aurora B和BubR1发挥调控纺锤体检验点的功能(8)。研究提示由于Chk1被抑制后会导致很多癌细胞死亡,Chk1可以作为癌症治疗的药物靶点(9)。

  1. Liu, Q. et al. (2000) Genes Dev 14, 1448-59.
  2. Zhao, H. and Piwnica-Worms, H. (2001) Mol Cell Biol 21, 4129-39.
  3. Jiang, K. et al. (2003) J Biol Chem 278, 25207-17.
  4. Martin, S.A. and Ouchi, T. (2008) Mol Cancer Ther 7, 2509-16.
  5. Chen, M.S. et al. (2003) Mol Cell Biol 23, 7488-97.
  6. Zeng, Y. et al. (1998) Nature 395, 507-10.
  7. Löffler, H. et al. (2006) Cell Cycle 5, 2543-7.
  8. Zachos, G. et al. (2007) Dev Cell 12, 247-60.
  9. Garber, K. (2005) J Natl Cancer Inst 97, 1026-8.

Application References

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For Research Use Only. Not For Use In Diagnostic Procedures.

U.S. Patent No. 5,675,063.

Cell Signaling Technology is a trademark of Cell Signaling Technology, Inc.

Cell Signaling Technology® is a trademark of Cell Signaling Technology, Inc.

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