| 货号 | 9917T |
| 描述 | The Cell Cycle/Checkpoint Antibody Sampler Kit provides a fast and economical means of evaluating multiple proteins involved in the cell cyle and checkpoint control. The kit contains enough primary and secondary antibody to perform four Western blot experiments.Cell Cycle/Checkpoint Antibody Sampler Kit能够快速经济地评价细胞周期和检验点控制的多个蛋白。该试剂盒内含足够4次western blot实验的一抗和二抗。 |
| 目标/特异性 | Phospho-cdc2 (Tyr15) (10A11) Rabbit mAb detects endogenous levels of cdc2 protein only when phosphorylated at tyrosine 15. Based on sequence similarity, the antibody may cross-react with CDK2 and CDK3. Phospho-Chk2 (Thr68) (C13C1) Rabbit mAb detects endogenous levels of Chk2 only when phosphorylated at Thr68. Phospho-Chk1 (Ser345) Antibody detects Chk1 only when phosphorylated at Ser345 and does not cross-react with other proteins. Phospho-Rb (Ser795) Antibody detects Rb only when phosphorylated at Ser795 and does not cross-react with Rb phosphorylated at other sites. Phospho-Rb (Ser807/811) (D20B12) XP® Rabbit mAb recognizes endogenous levels of Rb protein only when phosphorylated at Ser807, Ser811, or at both sites. This antibody does not cross-react with Rb phosphorylated at Ser608. Phospho-p53 (Ser15) (16G8) Mouse mAb detects endogenous levels of p53 only when phosphorylated at Ser15. The antibody does not cross-react with p53 phosphorylated at other sites. |
| 供应商 | CST |
| 背景 | The cell division cycle demands accuracy to avoid the accumulation of genetic damage. This process is controlled by molecular circuits called "checkpoints" that are common to all eukaryotic cells (1). Checkpoints monitor DNA integrity and cell growth prior to replication and division at the G1/S and G2/M transitions, respectively. The cdc2-cyclin B kinase is pivotal in regulating the G2/M transition (2,3). Cdc2 is phosphorylated at Thr14 and Tyr15 during G2-phase by the kinases Wee1 and Myt1, rendering it inactive. The tumor suppressor protein retinoblastoma (Rb) controls progression through the late G1 restriction point (R) and is a major regulator of the G1/S transition (4). During early and mid G1-phase, Rb binds to and represses the transcription factor E2F (5). The phosphorylation of Rb late in G1-phase by CDKs induces Rb to dissociate from E2F, permitting the transcription of S-phase-promoting genes. In vitro, Rb can be phosphorylated at multiple sites by cdc2, cdk2, and cdk4/6 (6-8). DNA damage triggers both the G2/M and the G1/S checkpoints. DNA damage activates the DNA-PK/ATM/ATR kinases, which phosphorylate Chk at Ser345 (9), Chk2 at Thr68 (10) and p53 (11). The Chk kinases inactivate cdc25 via phosphorylation at Ser216, blocking the activation of cdc2.细胞分裂周期需要准确性来避免遗传损伤积累。这个过程被常见于所有真核细胞的称为“检验点”的分子电路控制(1)。检验点分别先于复制和G1/S期以及G2/ M转换期分化监测DNA的完整性和细胞的生长。Cdc2-细胞周期蛋白B激酶在调节G2/M期转换中很关键(2,3)。在G2期Wee1和MYT1蛋白激酶介导的Cdc2 的Thr14和Tyr15磷酸化，使其处于非活动状态。肿瘤抑制蛋白视网膜母细胞瘤(Rb) 通过后期G1限制点(R)控制进展，而且是G1/ S过渡的重要调节器(4)。在G1期早期和中期，Rb结合并抑制转录因子E2F(5)。在G1期后期由CDKs磷酸化的Rb诱导Rb从E2F解离，允许S期促进基因的转录。在体外，Rb可以在多个位点被cdc2，cdk2和cdk4/6磷酸化(6-8)。DNA损伤能够引发G2/ M和G1/ S检验点。DNA损伤激活DNA-PK/ATM/ATR激酶，从而磷酸化Chk 的Ser345 (9)和Chk2的Thr68(10)以及p53(11)。CHK激酶能够通过Ser216磷酸化灭活cdc25，阻断cdc2的激活。 |
| 存放说明 | -20C |
| 参考文献 | Nurse, P. (1997) Cell 91, 865-7. Norbury, C. and Nurse, P. (1992) Annu Rev Biochem 61, 441-70. Watanabe, N. et al. (1995) EMBO J 14, 1878-91. Sherr, C.J. (1996) Science 274, 1672-7. Dyson, N. (1998) Genes Dev 12, 2245-62. Kitagawa, M. et al. (1996) EMBO J 15, 7060-9. Lundberg, A.S. and Weinberg, R.A. (1998) Mol Cell Biol 18, 753-61. Harbour, J.W. et al. (1999) Cell 98, 859-69. Zhao, H. and Piwnica-Worms, H. (2001) Mol Cell Biol 21, 4129-39. Matsuoka, S. et al. (2000) Proc Natl Acad Sci USA 97, 10389-94. Tibbetts, R.S. et al. (1999) Genes Dev 13, 152-7. |
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. | |
Flow cytometric analysis of Jurkat cells using Phospho-Rb (Ser807/811) (D20B12) XP® Rabbit mAb compared to Propidium Iodide (PI)/RNase Staining Solution #4087. | |
Immunohistochemical analysis of paraffin-embedded mouse spleen using Phospho-Rb (Ser807/811) (D20B12) XP® Rabbit mAb. | |
Western blot analysis of extracts from WI-38 cells, serum-starved for 3 days (-) or serum-starved for 3 days followed by treatment with 10% serum for 2 days (+), using Phospho-Rb (Ser807/811) (D20B12) XP® Rabbit mAb #8516. | |
Immunoprecipitation of phospho-Rb (Ser807/811) from MCF7 cell extracts using Phospho-Rb (Ser807/811) (D20B12) XP® Rabbit mAb (lane 2). Western blot was performed using the same antibody. Lane 1 is 10% input. | |
Western blot analysis of extracts from WI-38 cells, serum-starved for 3 days (-) or serum-starved for 3 days followed by treatment with 10% serum for 2 days (+), using Phospho-Rb (Ser807/811) (D20B12) XP® Rabbit mAb. | |
Western blot analysis of extracts from MCF7 cells, untreated (-) or treated with calf intestinal phosphatase (CIP) and λ phosphatase (+), using Phospho-Rb (Ser807/811) (D20B12) XP® Rabbit mAb (upper) or Rb (4H1) Mouse mAb #9309 (lower). | |
Immunohistochemical analysis of paraffin-embedded human lung carcinoma, control (left) or λ phosphatase-treated (right), using Phospho-Rb (Ser807/811) (D20B12) XP® Rabbit mAb. | |
Immunohistochemical analysis of paraffin-embedded human ovarian serous adenocarcinoma using Phospho-Rb (Ser807/811) (D20B12) XP® Rabbit mAb in the presence of control peptide (left) or antigen-specific peptide (right). | |
Immunohistochemical analysis of paraffin-embedded human colon carcinoma using Phospho-Rb (Ser807/811) (D20B12) XP® Rabbit mAb. | |
Confocal immunofluorescent analysis of MCF7 (left) and BT-549 (right) cells, untreated (upper) or λ phosphatase-treated (lower) using Phospho-Rb (Ser807/Ser811) (D20B12) XP® Rabbit mAb (green). Actin filaments were labeled with DY-554 phalloidin (red). Blue pseudocolor = DRAQ5® #4084 (fluorescent DNA dye). | |
Western blot analysis of extracts from MvILu cells treated with UV or hydroxyurea (20 mM) for the indicated times, using Phospho-p53 (Ser15) Antibody #9284.Western blot方法检测紫外或羟基脲(20 mM) 处理不同时间的MvILu细胞提取物，使用的抗体为Phospho-p53 (Ser15) Antibody #9284。 | |
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). | |
Immunohistochemical analysis of paraffin-embedded human lung carcinoma using Phospho-Chk2 (Thr68) (C13C1) Rabbit mAb. | |
Flow cytometric analysis of HeLa cells, untreated (blue) and UV-treated (green), using Phospho-Chk1 (Ser345) (133D3) Rabbit mAb. |
