Cell Signaling Technology

Product Pathways - Cell Cycle / Checkpoint

Phospho-Cyclin B1 (Ser147) Antibody #4131

b1   Cyclin   Cyclin B   cyclinb1  

No. Size Price
4131S 100 µl ( 10 western blots ) ¥3,900.00 现货查询 购买询价
4131 carrier free & custom formulation / quantityemail request
Applications Dilution Species-Reactivity Sensitivity MW (kDa) Isotype
W 1:1000 Human,Mouse,Rat, Endogenous 55 Rabbit

Species cross-reactivity is determined by western blot.

Applications Key: W=Western Blotting,

Homology

Species predicted to react based on 100% sequence homology: Pig,

Specificity / Sensitivity

Phospho-Cyclin B1 (Ser147) Antibody detects endogenous levels of cyclin B1 only when phosphorylated at serine 147. Phospho-Cyclin B1 (Ser147) Antibody能够检测内源性的丝氨酸(147位点)磷酸化的cyclin B1蛋白。

Source / Purification

Polyclonal antibodies are produced by immunizing animals with a synthetic phosphopeptide corresponding to residues surrounding Ser147 of human cyclin B1. Antibodies are purified by protein A and peptide affinity chromatography. 该单克隆抗体是由合成的人源的针对cyclin B1蛋白丝氨酸(147位)磷酸化肽段免疫动物,利用A蛋白和多肽亲和层析方法纯化生产的。

Western Blotting

Western Blotting

Western blot analysis of extracts from 293 cells, using Phospho-Cyclin B1 (Ser147) Antibody (A) and Cyclin B1 (V152) Monoclonal Antibody (C). Treatment of the membrane with calf intestinal alkaline phosphatase (CIP) after Western transfer abolishes the phospho-cyclin B1 signal(B), but has no effect on the total cyclin B1 signal (D). western blot方法检测293细胞提取物,使用的抗体为 Phospho-Cyclin B1 (Ser147) Antibody (A) 和Cyclin B1 (V152) Monoclonal Antibody (C)。转膜后用牛小肠碱性磷酸酶(CIP)处理western膜,可以消除phospho-cyclin B1 信号(B),但是对于 cyclin B1 总蛋白没有影响。

Western Blotting

Western Blotting

Western blot analysis of extracts from asynchronous cells (lane 1), S-phase and G2/M blocked cells (lane 2), or synchronized cells collected at various time points following release from this block (lanes 3-7) using Phospho-Cyclin B1 (Ser147) Antibody (upper). Cell cycle synchronization was verified by flow cytometric analysis of DNA content (lower). (Data kindly provided by Ethan Kohn and Alan Eastman, Dartmouth Medical School, Hanover, NH). Western blot方法检测异步细胞(泳道1),S期和G2/M期阻滞细胞(泳道2),或解除该阻滞后于不同时间点收集的同步细胞(泳道3-7),使用的抗体为Phospho-Cyclin B1 (Ser147) Antibody (上图). 采用流式细胞术检测DNA含量用以证明细胞周期同步(下图)。(数据由达特茅斯医学院的 Ethan Kohn 和Alan Eastman 提供)

Western Blotting

Western Blotting

Western blot analysis of extracts from HT29 cells, untreated (cont.) or treated with 50 ng/ml nocodazole (M), using Phospho-cyclin B1 (Ser147) Antibody (upper), Cyclin B1 (V152) Monoclonal Antibody (middle), or cdc25C Antibody #9522 (lower). western blot方法检测HT29细胞提取物:未处理(cont.) 或nocodazole处理(50 ng/ml) 处理(M)。使用的抗体为Phospho-cyclin B1 (Ser147) Antibody (上图), Cyclin B1 (V152) Mouse mAb (中图), 或Cdc25C Antibody #9522 (下图).

Background

Entry of all eukaryotic cells into mitosis is regulated by activation of cdc2 kinase. Activation of cdc2 is controlled at several steps including cyclin B1 binding, phosphorylation of cdc2 at Thr161 and dephosphorylation of cdc2 at Thr14/Tyr15 (1-5). The protein levels of CDK inhibitors and the CDK-associated cyclins are regulated by phosphorylation, ubiquitination and degradation, allowing for a stoichiometric regulation of cell cycle events (6). Four cyclin B1 phosphorylation sites (Ser126, 128, 133 and 147) are located in the cytoplasmic retention signal (CRS) domain and are thought to regulate the translocation of cyclin B1 to the nucleus at the G2/M checkpoint (8-10). Phosphorylation of cyclin B1 is required for cdc25C-dependent dephosphorylation of Tyr15 within cdc2 and subsequent cdc2/cyclin B1 activation (11). While cdc2/cyclin B1 itself can phosphorylate Ser126 and Ser128 (8), polo-like kinase 1 phosphorylates cyclin B1 preferentially at Ser133 and possibly at Ser147 as well (11-13). cdc2蛋白激酶的激活参与调节所有的真核细胞进入有丝分裂。cdc2的激活受多个步骤的调控,包括周期蛋白B的结合、cdc2苏氨酸161位的磷酸化和苏氨酸14位和15位的去磷酸化(1-5)。CDK抑制剂和CDK相关的细胞周期蛋白的水平可通过磷酸化、泛素化和降解来调节,允许了细胞周期事件的化学计量调节(6)。四个周期蛋白B1磷酸化位点(丝氨酸126位,128位,133位和147位)位于胞质滞留信号(CRS)结构域,并可以调节G2/S检验点周期蛋白B1的核迁移(8-10)。周期蛋白B1的磷酸化参与了cdc25C依赖的cdc2 酪氨酸15位的去磷酸化和随后的cdc2/周期蛋白B1 激活(11)。尽管cdc2/周期蛋白B1本身能够磷酸化丝氨酸126位和128位(8),polo样蛋白激酶优先磷酸化丝氨酸133位,也可能磷酸化丝氨酸147位(11-13)。

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