| 货号 | 9870T |
| 描述 | The Cell Cycle Regulation Sampler Kit II provides an economical means of evaluating cell cycle proteins. The kit contains enough primary and secondary antibodies to perform four western blot experiments.Cell Cycle Regulation Sampler Kit II能够经济地评价细胞周期蛋白。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 | Each antibody in the Cell Cycle Regulation Sampler Kit II detects endogenous levels of its target protein and does not typically cross react with other family members. Cyclin B1 (D5C10) XP® Rabbit mAb recognizes endogenous levels of total cyclin B1 protein. This antibody also detects a 100 kDa protein of unknown origin in some cell lines. Activation state antibodies recognize target proteins only when phosphorylated at the indicated residue. 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. |
| 供应商 | CST |
| 背景 | The critical regulatory step in activating cdc2 during progression into mitosis appears to be dephosphorylation of cdc2 at Tyr15 and Thr14 (1). Phosphorylation of cdc2 by the protein kinases Wee1 and Myt1 at Thr14 and Tyr15 results in inhibition of cdc2 (2,3). Progression through the G1/S checkpoint and initiation of DNA replication requires cyclin E; traversing the G2/M checkpoint to initiate mitosis requires cyclin B, and cyclin A may be required for both S-phase and M-phase (4). The versatile p21 cyclin-dependent kinase inhibitor, which interacts with several cyclin-CDK complexes to regulate cyclin-CDK during the cell cycle, is regulated by phosphorylation and ubiquitin-mediated degradation (5). Phosphorylation of histone H3 at Ser10 and neighboring residues correlates with chromosomal condensation, which is essential for segregation of chromosomes during mitosis (6).激活细胞周期进程中的cdc2进入有丝分裂的关键调节步骤似乎是cdc2 的Tyr15和Thr14去磷酸化(1)。蛋白激酶Wee1和Myt1介导的cdc2的Thr14和Tyr15磷酸化将导致cdc2抑制(2,3)。通过G1/S检验点的进程和DNA复制起始需要细胞周期蛋白E；穿越G2/ M检验点开始有丝分裂需要细胞周期蛋白B，而细胞周期S期和M期可能都需要周期蛋白A(4)。多功能的p21细胞周期蛋白依赖性激酶抑制剂，它与几个细胞周期蛋白-CDK复合物相互作用来调节在细胞周期中的细胞周期蛋白-CDK，接受磷酸化和泛素介导降解的调节(5)。组蛋白H3的Ser10和邻近残基的磷酸化与染色体缩合相关，这是有丝分裂染色体分离必不可少的(6)。Application References |
| 存放说明 | -20C |
| 参考文献 | 1 . Norbury, C. et al. (1991) EMBO J 10, 3321-9. 2 . Wells, N.J. et al. (1999) J Cell Sci 112 ( Pt 19), 3361-71. 3 . McGowan, C.H. and Russell, P. (1993) EMBO J 12, 75-85. 4 . Pagano, M. et al. (1992) EMBO J 11, 961-71. 5 . Abbas, T. and Dutta, A. (2009) Nat Rev Cancer 9, 400-14. 6 . Hendzel, M.J. et al. (1997) Chromosoma 106, 348-60. |
Flow cytometric analysis of Jurkat cells using Cyclin B1 (D5C10) XP® Rabbit mAb and Propidium Iodide (PI)/RNase Staining Solution #4087 (DNA content). Anti-rabbit IgG (H+L), F(ab)2 Fragment (Alexa Fluor® 488 Conjugate) #4412 was used as a secondary antibody. | |
Western blot analysis of extracts from HT-29 cells, synchronized in S-phase by double thymidine block (2 nM, 16 hr) followed by release into fresh media for the indicated time, using Cyclin B1 (D5C10) XP® Rabbit mAb (upper) or β-Actin (D6A8) Rabbit mAb #8457 (lower). | |
Western blot analysis of extracts from various cell lines using Cyclin B1 (D5C10) XP® Rabbit mAb. | |
Confocal immunofluorescent analysis of HT-29 cells using Cyclin B1 (D5C10) XP® Rabbit mAb (green). Blue pseudocolor = DRAQ5®#4084 (fluorescent DNA dye). | |
Western blot analysis of extracts from HeLa cells, transfected with 100 nM SignalSilence® Control siRNA (Fluorescein Conjugate) #6201 (-) or SignalSilence® p21 Waf1/Cip1 siRNA II (+), using p21 Waf1/Cip1 (12D1) Rabbit mAb #2947 and α-Tubulin (11H10) Rabbit mAb #2125. The p21 Waf1/Cip1 (12D1) Rabbit mAb confirms silencing of p21 Waf1/Cip1 expression and α-Tubulin (11H10) Rabbit mAb is used to control for loading and specificity of p21 Waf1/Cip1 siRNA. | |
Western blot analysis of extracts from various cell types using p21 Waf1/Cip1 (12D1) Rabbit mAb #2947.Western blot 方法检测不同细胞提取物，使用的抗体为p21 Waf1/Cip1 (12D1) Rabbit mAb #2947。 | |
Western analysis of extracts from HeLa cells, untreated or treated with doxorubicin (0.5 μM, 24 hours) or with nocodazole (50 ng/ml, 24 hours), using Cyclin A (BF683) Mouse mAb #4656.Western blot方法检测未处理、阿霉素(0.5 μM)处理24小时或诺考达唑(50 ng/ml)处理24小时 的Hela细胞，使用的抗体为Cyclin A (BF683) Mouse mAb #4656。 | |
Western blot analysis of extracts from HT-29 cells, untreated (lane 1), nocodazole-treated (50 ng/ml, lane 2), or UV-treated (lane 3), using Myt1 Antibody #4282.Western blot 检测HT-29细胞提取物，未处理组（泳道1），诺考达唑处理组（50 ng/ml，泳道2）或紫外处理组（泳道3），使用的抗体为Myt1 Antibody #4282。 | |
Western blot analysis of extracts from HeLa cells, untreated or treated with nocodazole (0.1 mg/ml for 18 hours), using Phospho-Histone H3 (Ser10) (D2C8) XP® Rabbit mAb #3377 (upper) or Histone H3 Antibody #9715 (lower). Phospho-specificity of the antibody is shown by further treatment of the lysate with λ phosphatase.Western blot 方法检测未处理或诺考达唑 (0.1 mg/ml)处理18小时的Hela细胞, 使用的抗体为Phospho-Histone H3 (Ser10) (D2C8) XP® Rabbit mAb #3377 (上图) 或 Histone H3 Antibody #9715 (下图)。抗体的磷酸化特异性使用λ磷酸酶处理的裂解液检测。 | |
Western blot analysis of extracts from MCF7, SK-N-MC and HeLa cells, untreated or treated with the proteasome inhibitor MG-132, using Cyclin E2 Antibody #4132.Western blot方法检测未处理或蛋白酶抑制剂MG-132处理的MCF7\SK-N-MC和HeLa细胞提取物，使用的抗体为Cyclin E2 Antibody #4132。 | |
Western blot analysis of extracts from various cell types using Cyclin B1 Antibody #4138.Western blot 方法检测不同细胞提取物，使用的抗体为Cyclin B1 Antibody #4138。 | |
Western blot analysis of extracts from Saos-2 cells, untreated or treated with hydroxyurea or nocodazole, using Phospho-cdc2 (Tyr15) Antibody #9111 (upper) or cdc2 Antibody #9112 (lower).Western blot方法检测未处理或羟基脲、诺考达唑处理的Saos-2细胞提取物，使用的抗体为 Phospho-cdc2 (Tyr15) Antibody #9111 (上图) 或 cdc2 Antibody #9112 (下图)。 | |
Western blot analysis of extracts from A431 and H441 cells, untreated or EGF-treated, using Phospho-Wee1 (Ser642) (D47G5) Rabbit mAb #4910 (upper) or Wee1 Antibody #4936 (lower).Western blot 方法检测未处理或EGF处理的A431和H441细胞提取物，使用的抗体为 Phospho-Wee1 (Ser642) (D47G5) Rabbit mAb #4910 (上图) 或 Wee1 Antibody #4936 (下图). | |
Flow cytometric analysis of untreated Jurkat cells, using Cyclin A2 (BF683) Mouse mAb versus propidium iodide (DNA content). Note decrease in expression of Cyclin A during late mitosis (arrow). | |
Western blot analysis of extracts from HeLa cells, either untreated or treated with nocodazole (100 ng/ml for 18 hours), using Phospho-Histone H3 (Ser10) (D2C8) XP® Rabbit mAb #3377 (upper) or Histone H3 Antibody #9715 (lower). Phospho-specificity of the antibody is shown by further treatment of the lysate with λ phosphatase. |
