Cell Signaling Technology

Product Pathways - Cell Cycle / Checkpoint

Phospho-p57 Kip2 (Thr310) Antibody #2558

CDKN1C   CDNC   KIP2   p57Kip2  

No. Size Price
2558S 100 µl ( 10 western blots ) ¥3,900.00 现货查询 购买询价
2558 carrier free & custom formulation / quantityemail request
Applications Dilution Species-Reactivity Sensitivity MW (kDa) Isotype
W 1:1000 Human, Endogenous 57 Rabbit
IF-IC 1:100

Species cross-reactivity is determined by western blot.

Applications Key: W=Western Blotting, IF-IC=Immunofluorescence (Immunocytochemistry),

Specificity / Sensitivity

Phospho-p57 Kip2 (Thr310) Antibody detects endogenous levels of p57 Kip2 only when phosphorylated at threonine 310. This antibody may cross-react with p27 Kip1 when phosphorylated at Thr187. Phospho-p57 Kip2 (Thr310) Antibody能够检测内源性苏氨酸(310位)磷酸化的p57 Kip2蛋白。该抗体也许与苏氨酸(187位)磷酸化的p27 Kip1有交叉反应。

Source / Purification

Polyclonal antibodies are produced by immunizing animals with a synthetic phosphopeptide corresponding to residues surrounding Thr310 of human p57 Kip2. Antibodies are purified by protein A and peptide affinity chromatography.该多克隆抗体是由合成的人源的针对p57 Kip2蛋白苏氨酸(310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。

Western Blotting

Western Blotting

Western blot analysis of extracts from untreated and dexamethasone-treated (50 nM, 16 h) HeLa cells, using Phospho-p57 Kip2 (Thr310) Antibody (upper) and p57 Kip2 Antibody #2557 (lower). Lysates were treated with lambda phosphatase NEB#P0753 (10,000 U/ml for 30 mins) to confirm the phospho-specificity of the phospho-antibody.采用Western blot 方法检测未处理和地塞米松(50 nM) 单独处理16小时的Hela细胞,使用的抗体为Phospho-p57 Kip2 (Thr310) Antibody (上图) 和 p57 Kip2 Antibody #2557 (下图) 。裂解液用 lambda 磷酸酶(10,000 U/ml)处理30分钟以确定磷酸化抗体的磷酸化特异性,磷酸酶的产品信息为NEB#P0753。

IF-IC

IF-IC

Confocal immunofluorescent analysis of HeLa cells, dexamethasone-treated (left) or phosphatase-treated (right), labeled with Phospho-p57 Kip2 (Thr310) Antibody (green). Actin filaments have been labeled with Alexa Fluor® 555 phalloidin (red).激光共聚焦免疫荧光方法检测Hela细胞,地塞米松处理组(左图)和磷酸酶处理组(右图), 标记抗体为Phospho-p57 Kip2 (Thr310) Antibody ,呈绿色。肌动蛋白纤维由Alexa Fluor® 555鬼笔环肽标记,呈红色。

Background

p27 Kip1 is a member of the Cip/Kip family of cyclin-dependent kinase inhibitors. Like its relatives, p57 Kip2 and p21 Waf1/Cip1, the ability to enforce the G1 restriction point is derived from its inhibitory binding to CDK2/cyclin E and other CDK/cyclin complexes. Expression levels of p27 are upregulated in quiescent cells and in cells treated with cAMP or other negative cell cycle regulators. Downregulation of p27 can be induced by treatment with interleukin 2 or other mitogens; this involves phosphorylation of p27 and its degradation by the ubiquitin-proteasome pathway (1-4).p27 Kip1是周期蛋白依赖性激酶抑制剂Cip/Kip家族的一员。同其类似物一样,如p57 Kip2 和p21 Waf1/Cip1,其巩固G1限制点的能力源于CDK2/cyclin E和其它CDK/cyclin复合体的结合抑制。在静止期细胞和cAMP或其它细胞周期阴性调节剂处理的细胞中,p27的表达是上调的。白细胞介素2或者其它有丝分裂原能够诱导p27的下调;该过程涉及p27的磷酸化和泛素化-蛋白酶体途径介导的降解(1-4)。

Levels of p57 Kip2 are controlled by ubiquitination/degradation via the Skp1/Cul1/F-box-type E3 ubiquitin ligase complex SCF-Skp2, and this effect is dependent on Thr310 (5). A similar threonine phosphorylation site in p27 Kip1, Thr187, has also been shown to regulate protein stability (6).p57 Kip2表达水平通过Skp1/Cul1/F-box-type E3 泛素连接酶复合物 SCF-Skp2被.泛素化/降解途径控制。这一效应依赖于苏氨酸(310位)(5)。p27 Kip1中对应的苏氨酸磷酸化位点为苏氨酸(187位),同样具有调节蛋白稳定性的作用(6)。

  1. Lloyd, R.V. et al. (1999) Am J Pathol 154, 313-23.
  2. Polyak, K. et al. (1994) Genes Dev 8, 9-22.
  3. Kato, J.Y. et al. (1994) Cell 79, 487-96.
  4. Vlach, J. et al. (1997) EMBO J 16, 5334-44.
  5. Kamura, T. et al. (2003) Proc. Natl. Acad. Sci. USA 100, 10231-10236.
  6. Ishida, N. et al. (2000) J. Biol. Chem. 275, 25146-25154.

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Cell Signaling Technology is a trademark of Cell Signaling Technology, Inc.

Alexa Fluor is a registered trademark of Life Technologies Corporation.

Cell Signaling Technology® is a trademark of Cell Signaling Technology, Inc.

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