Cell Cycle/Checkpoint Antibody Sampler Kit

• 产品编号：CST-9917S      品牌：Cell Signaling Technology       原厂货号：9917S
• 产品分类：抗体 > 一抗 > 复合抗体试剂盒
• 应用分类：

其它组分：

描述：

.Phospho-cdc2 (Tyr15) Antibody能够检测酪氨酸（15位）磷酸化的cdc2和 cdk2 蛋白，且不与cdk4, cdk6 或cdk7发生交叉反应。Phospho-Chk2 (Thr68) Antibody 能够检测苏氨酸（68位）磷酸化的Chk2蛋白，且不与非磷酸化Chk2发生交叉反应。Phospho-Chk1 (Ser345) Antibody能够检测丝氨酸（345位）磷酸化的Chk1 蛋白，且不与其他蛋白发生交叉反应。Phospho-Rb (Ser795) 和 (Ser807/811) Antibodies 能够检测特定位点磷酸化的Rb蛋白，且不与其他位点发生磷酸化的Rb蛋白发生交叉反应。Phospho-p53 (Ser15) Antibody 能够检测丝氨酸（15位）磷酸化的p53 蛋白且不与非磷酸化p53蛋白发生交叉反应。多克隆抗体分别由合成的人源的针对cdc2蛋白酪氨酸（15位）的磷酸化肽段，Chk1蛋白丝氨酸（345位）的磷酸化肽段，Chk2蛋白苏氨酸（68位）的磷酸化肽段，Rb蛋白丝氨酸（795、807/811位）的磷酸化肽段免疫动物，采用蛋白A和多肽亲和层析技术纯化生产的。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。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。

1. Nurse, P. (1997) Cell 91, 865-7.
2. Norbury, C. and Nurse, P. (1992) Annu Rev Biochem 61, 441-70.
3. Watanabe, N. et al. (1995) EMBO J 14, 1878-91.
4. Sherr, C.J. (1996) Science 274, 1672-7.
5. Dyson, N. (1998) Genes Dev 12, 2245-62.
6. Kitagawa, M. et al. (1996) EMBO J 15, 7060-9.
7. Lundberg, A.S. and Weinberg, R.A. (1998) Mol Cell Biol 18, 753-61.
8. Harbour, J.W. et al. (1999) Cell 98, 859-69.
9. Zhao, H. and Piwnica-Worms, H. (2001) Mol Cell Biol 21, 4129-39.
10. Matsuoka, S. et al. (2000) Proc Natl Acad Sci USA 97, 10389-94.
11. Tibbetts, R.S. et al. (1999) Genes Dev 13, 152-7.

原厂资料：

Specificity / Sensitivity

Phospho-cdc2 (Tyr15) Antibody detects cdc2 and cdk2 only when phosphorylated at Tyr15 and does not cross-react with cdk4, cdk6 or cdk7. Phospho-Chk2 (Thr68) Antibody detects Chk2 only when phosphorylated at Thr68 and does not react with nonphosphorylated Chk2. Phospho-Chk1 (Ser345) Antibody detects Chk1 only when phosphorylated at Ser345 and does not cross-react with other proteins. Phospho-Rb (Ser795) and (Ser807/811) Antibodies detect Rb only when phosphorylated at the indicated sites and do not cross-react with Rb phosphorylated at other sites. Phospho-p53 (Ser15) Antibody detects p53 only when phosphorylated at Ser15 and does not react with nonphosphorylated p53.

Source / Purification

Polyclonal antibodies are produced by immunizing animals with synthetic phosphopeptides corresponding to residues surrounding Tyr15 of human cdc2; Ser345 of human Chk1; Thr68 of human Chk2; Ser795 and Ser807/811 of human Rb; and Ser15 of human p53. Polyclonal antibodies are purified by protein A and peptide affinity chromatography.

Description

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.

Background

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.

1. Nurse, P. (1997) Cell 91, 865-7.
2. Norbury, C. and Nurse, P. (1992) Annu Rev Biochem 61, 441-70.
3. Watanabe, N. et al. (1995) EMBO J 14, 1878-91.
4. Sherr, C.J. (1996) Science 274, 1672-7.
5. Dyson, N. (1998) Genes Dev 12, 2245-62.
6. Kitagawa, M. et al. (1996) EMBO J 15, 7060-9.
7. Lundberg, A.S. and Weinberg, R.A. (1998) Mol Cell Biol 18, 753-61.
8. Harbour, J.W. et al. (1999) Cell 98, 859-69.
9. Zhao, H. and Piwnica-Worms, H. (2001) Mol Cell Biol 21, 4129-39.
10. Matsuoka, S. et al. (2000) Proc Natl Acad Sci USA 97, 10389-94.
11. Tibbetts, R.S. et al. (1999) Genes Dev 13, 152-7.

注意事项：

Storage: Supplied in 10 mM sodium HEPES (pH 7.5), 150 mM
NaCl, 100 µg/ml BSA and 50% glycerol. Store at –20°C.
Do not aliquot the antibodies.