| 货号 | 9931T |
| 描述 | The Phospho-Chk1/2 Antibody Sampler Kit offers an economical means to evaluate the phosphorylation status of Chk1 and Chk2 on multiple residues. The kit contains enough primary and secondary antibodies to perform four Western blot experiments with each primary antibody.Phospho-Chk1/2 Antibody Sampler Kit能够经济地评价Chk1和Chk2蛋白在多位点的磷酸化状态。该试剂盒包含足够四次western blot所需的一抗和二抗。 |
| 目标/特异性 | Each antibody in the Phospho-Chk1/2 Antibody Sampler Kit detects endogenous levels of its respective target protein. |
| 供应商 | CST |
| 背景 | 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 reentry 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). Chk1 has emerged as a drug target for cancer therapy as its inhibition leads to cell death in many cancer cell lines (9).Chk1激酶充当ATM/ATR激酶的下游，在DNA损伤检验点的控制、胚胎发育和肿瘤抑制中扮演着重要角色(1)。Chk1的激活涉及Ser317和Ser345的磷酸化，其发生响应于DNA复制中断和某些形式的基因毒性应激(2)。Ser345的磷酸化有助于检验点激活后Chk1定位到细胞核(3)，而Ser317的磷酸化随伴PTEN的位点特异性磷酸化并允许DNA复制停滞后再次进入细胞周期(4)。Chk1的细胞周期检验点机制的发挥部分通过调节磷酸酶的cdc25家族。Chk1磷酸化cdc25A靶向其蛋白水解并通过14-3-3结合抑制其活性(5)。活化的Chk1可以通过磷酸化Ser216灭活cdc25C，阻断cdc2的激活有丝分裂期的转换(6)。中心体Chk1已被证明能够磷酸化cdc25B和抑制其CDK1细胞周期素B1的激活，从而废除有丝分裂纺锤体的形成和染色质聚集(7)。此外，通过调节极光激酶B和BubR1，Chk1在纺锤体检验点的功能中起到了重要作用。由于Chk1的抑制作用可导致许多肿瘤细胞株死亡，它已成为用于癌症治疗的药物靶标(9)。 |
| 存放说明 | -20C |
| 参考文献 | Liu, Q. et al. (2000) Genes Dev 14, 1448-59. Zhao, H. and Piwnica-Worms, H. (2001) Mol Cell Biol 21, 4129-39. Jiang, K. et al. (2003) J Biol Chem 278, 25207-17. Martin, S.A. and Ouchi, T. (2008) Mol Cancer Ther 7, 2509-16. Chen, M.S. et al. (2003) Mol Cell Biol 23, 7488-97. Zeng, Y. et al. (1998) Nature 395, 507-10. Löffler, H. et al. (2006) Cell Cycle 5, 2543-7. Zachos, G. et al. (2007) Dev Cell 12, 247-60. Garber, K. (2005) J Natl Cancer Inst 97, 1026-8. Allen, J. B. et al. (1994) Genes Dev. 8, 2401-2415. Weinert, T. A. et al. (1994) Genes Dev. 8, 652-665. Murakami, H. and Okayama, H. (1995) Nature 374, 817-819. Kastan, M.B. and Lim, D.S. (2000) Nat. Rev. Mol. Cell Biol. 1, 179-186. Matsuoka, S. et al. (2000) Proc. Natl. Acad. Sci. USA 97, 10389-10394. Melchionna, R. et al. (2000) Nat. Cell Biol. 2, 762-765. Ahn, J. Y. et al. (2000) Cancer Res. 60, 5934-5936. |
Western blot analysis of extracts from various cell lines using Chk1 (2G1D5) Mouse mAb #2360. | |
Immunoprecipitation of phospho-Chk1 (Ser317) from 293 cell extracts treated with UV (100 mJ, 1 hr recovery) using Phospho-Chk1 (Ser317) (D12H3) XP® Rabbit mAb (lane 2) or Rabbit (D1AG) mAb IgG XP®Isotype Control #3900 (lane 3). Lane 1 is 10% input. | |
Western blot analysis of extracts from 293 and NIH/3T3 cells, untreated (-) or UV-treated (100 mJ, 1 hr recovery; +), using Phospho-Chk1 (Ser317) (D12H3) XP® Rabbit mAb. The blot on the right was treated with calf intestinal phosphatase (CIP) before western blot. | |
Confocal immunofluorescent analysis of HeLa cells, untreated (left), UV-treated (center), or UV and λ phosphatase-treated (right), using Phospho-Chk1 (Ser317) (D12H3) XP® Rabbit mAb (green). Actin filaments were labeled with DY-554 phalloidin (red). | |
Immunohistochemical analysis of paraffin-embedded human ovarian serous adenocarcinoma using Chk2 (D9C6) XP® Rabbit mAb. | |
Immunohistochemical analysis of paraffin-embedded human lung carcioma using Chk2 (D9C6) XP® 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. | |
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). | |
Immunohistochemical analysis of paraffin-embedded cell pellets, HCT 116 (left) or MDA-MB-231 (right), using Chk2 (D9C6) XP® Rabbit mAb. | |
Western blot analysis of extracts from various cell lines using Chk2 (D9C6) XP® Rabbit mAb. | |
Western blot analysis of extracts from HeLa cells, transfected with 100 nM SignalSilence® Control siRNA (Fluorescein Conjugate) #6201 (-) or SignalSilence® Chk1 siRNA I #6241 or SignalSilence® Chk1 siRNA II (+), using Chk1 (2G1D5) Mouse mAb #2360 and β-Actin (13E5) Rabbit mAb #4970. Chk1 (2G1D5) Mouse mAb confirms silencing of Chk1 expression and β-Actin (13E5) Rabbit mAb is used to control for loading and specificity of Chk1 siRNA. | |
Western blot analysis of extracts from 293 cells, untreated or UV-treated (50 mJ/cm2, 2hrs), using Phospho-Chk2 (Ser516) Antibody #2669 (upper) or Chk2 Antibody #2662 (lower).Western blot 方法检测未处理和紫外处理(50 mJ/cm2, 2小时)的293细胞提取物，使用的抗体为 Phospho-Chk2 (Ser516) Antibody #2669 (上图) 或Chk2 Antibody #2662 (下图). | |
Western blot analysis of extracts from HeLa, COS, NIH/3T3 and C6 cells, untreated or UV-treated, using Phospho-Chk1 (Ser345) (133D30) Rabbit mAb #2348.Western blot 方法检测未处理和紫外处理的HeLa, COS, NIH/3T3 和C6 细胞，使用的抗体为Phospho-Chk1 (Ser345) (133D30) Rabbit mAb #2348。 | |
Confocal immunofluorescent analysis of C2C12 cells, untreated (left) or UV-treated (right), using Phospho-Chk1 (Ser345) (133D3) Rabbit mAb (green). Actin filaments have been labeled with DY-554 phalloidin (red). | |
Flow cytometric analysis of HeLa cells, untreated (blue) and UV-treated (green), using Phospho-Chk2 (Thr68) Antibody. |
