Revision 2

#9555Store at -20C

1 个试剂盒

(3 x 20 microliters)

Cell Signaling Technology

Orders: 877-616-CELL (2355) [email protected]

Support: 877-678-TECH (8324)

Web: [email protected] cellsignal.com

3 Trask LaneDanversMassachusetts01923USA
For Research Use Only. Not for Use in Diagnostic Procedures.
Product Includes Product # Quantity Mol. Wt Isotype/Source
Phospho-cdc25C (Thr48) Antibody 9527 20 µl 75 kDa Rabbit 
Phospho-cdc25C (Ser216) (63F9) Rabbit mAb 4901 20 µl 60 kDa Rabbit IgG
cdc25C (5H9) Rabbit mAb 4688 20 µl 60, 75 kDa Rabbit IgG
Anti-rabbit IgG, HRP-linked Antibody 7074 100 µl Goat 

Please visit cellsignal.com for individual component applications, species cross-reactivity, dilutions, protocols, and additional product information.

Description

The cdc25C Antibody Sampler Kit provides an economical means to investigate the entry of eukaryotic cells into mitosis. The kit contains enough primary and secondary antibodies to perform two Western blots with each antibody.

Storage

Supplied in 10 mM sodium HEPES (pH 7.5), 150 mM NaCl, 100 µg/ml BSA, 50% glycerol and less than 0.02% sodium azide. Store at –20°C. Do not aliquot the antibody.

Background

Cdc25 is a protein phosphatase responsible for dephosphorylating and activating cdc2, a crucial step in regulating the entry of all eukaryotic cells into mitosis (1). cdc25C is constitutively phosphorylated at Ser216 throughout interphase by c-TAK1, while phosphorylation at this site is DNA damage-dependent at the G2/M checkpoint (2). When phosphorylated at Ser216, cdc25C binds to members of the 14-3-3 family of proteins, sequestering cdc25C in the cytoplasm and thereby preventing premature mitosis (3). The checkpoint kinases Chk1 and Chk2 phosphorylate cdc25C at Ser216 in response to DNA damage (4,5).
Full activation of cdc25C involves phosphorylation at more than 12 different sites by cdc2/cyclin B and Polo-like kinase, and the activity of Pin1, a peptidyl-prolyl isomerase (PPI) (6,7). Pin1 contains a WW domain that binds phospho-Ser/Thr-Pro sites and a catalytic PPI region that induces a cis/trans isomerization on phospho-Ser/Thr-Pro bonds (8). Thr48 and Thr67 of cdc25C interact directly with the WW domain of Pin1 when these sites are phosphorylated (9). Thr48 phosphorylation also mediates binding to CKS/p13SUC1 (10).

  1. Jessus, C. and Ozon, R. (1995) Prog. Cell Cycle Res. 1, 215-228.
  2. Peng, C.Y. et al. (1997) Science 277, 1501-1505.
  3. Kumagai, A. and Dunphy, W.G. (1999) Genes Dev. 13, 1067-1072.
  4. Blasina, A. et al. (1999) Curr. Biol. 9, 1-10.
  5. Furnari, B. et al. (1999) Mol. Biol. Cell 10, 833-845.
  6. Izumi, T. and Maller, J.L. (1993) Mol Biol Cell 4, 1337-50.
  7. Stukenberg, P.T. and Kirschner, M.W. (2001) Mol Cell 7, 1071-83.
  8. Yaffe, M.B. et al. (1997) Science 278, 1957-60.
  9. Lu, P.J. et al. (1999) Science 283, 1325-8.
  10. Landrieu, I. et al. (2001) J Biol Chem 276, 1434-8.

Background References

    Trademarks and Patents

    Cell Signaling Technology is a trademark of Cell Signaling Technology, Inc.
    All other trademarks are the property of their respective owners. Visit cellsignal.com/trademarks for more information.

    限制使用

    除非 CST 的合法授书代表以书面形式书行明确同意,否书以下条款适用于 CST、其关书方或分书商提供的书品。 任何书充本条款或与本条款不同的客书条款和条件,除非书 CST 的合法授书代表以书面形式书独接受, 否书均被拒书,并且无效。

    专品专有“专供研究使用”的专专或专似的专专声明, 且未专得美国食品和专品管理局或其他外国或国内专管机专专专任何用途的批准、准专或专可。客专不得将任何专品用于任何专断或治专目的, 或以任何不符合专专声明的方式使用专品。CST 专售或专可的专品提供专作专最专用专的客专,且专用于研专用途。将专品用于专断、专防或治专目的, 或专专售(专独或作专专成)或其他商专目的而专专专品,均需要 CST 的专独专可。客专:(a) 不得专独或与其他材料专合向任何第三方出售、专可、 出借、捐专或以其他方式专专或提供任何专品,或使用专品制造任何商专专品,(b) 不得复制、修改、逆向工程、反专专、 反专专专品或以其他方式专专专专专品的基专专专或技专,或使用专品开专任何与 CST 的专品或服专专争的专品或服专, (c) 不得更改或专除专品上的任何商专、商品名称、徽专、专利或版专声明或专专,(d) 只能根据 CST 的专品专售条款和任何适用文档使用专品, (e) 专遵守客专与专品一起使用的任何第三方专品或服专的任何专可、服专条款或专似专专

    Revision 2
    #9555

    cdc25C Antibody Sampler Kit

    cdc25C Antibody Sampler Kit: Image 1 Expand Image
    使用 cdc25C (5H9) Rabbit mAb 对 HeLa、293 和 K562 细胞提取物进行蛋白质印迹分析。
    cdc25C Antibody Sampler Kit: Image 2 Expand Image
    使用 Phospho-cdc25C (Ser216) Antibody(上)和 cdc25C (5H9) Rabbit mAb #4688(下),对未经处理、经 nocodazole 处理和 λ 磷酸酶处理的 HT29 细胞提取物进行蛋白质印迹分析。
    cdc25C Antibody Sampler Kit: Image 3 Expand Image
    一抗与靶标蛋白结合之后,与偶联 HRP 的二抗形成复合体。添加 LumiGLO®,在酶催化分解期间发光。
    cdc25C Antibody Sampler Kit: Image 4 Expand Image
    使用 Phospho-cdc25C (Thr48)(上)和 cdc25C (5H9) Rabbit mAb #4688(下),对未经处理、经 nocodazole 处理和 λ 磷酸酶处理的 HT29 细胞提取物进行蛋白质印迹分析。
    cdc25C Antibody Sampler Kit: Image 5 Expand Image
    使用 Phospho-cdc25C (Thr48) #9527(上)和 cdc25C (5H9) Rabbit mAb(下),对未经处理、经 nocodazole 处理和经 λ 磷酸酶处理的 HT29 细胞提取物进行蛋白质印迹分析。
    cdc25C Antibody Sampler Kit: Image 6 Expand Image
    使用 Phospho-cdc25C (Ser216) (63F9) Rabbit mAb(上)或 cdc25C Antibody #9522(下),对非磷酸化的或在体外被 Chk2 磷酸化的 GST-cdc25C 进行蛋白质印迹分析。
    cdc25C Antibody Sampler Kit: Image 7 Expand Image
    使用 Phospho-cdc25C (Ser216) (63F9) Rabbit mAb 对石蜡包埋的人乳腺癌进行免疫组织化学分析。
    cdc25C Antibody Sampler Kit: Image 8 Expand Image
    使用 Phospho-cdc25C (Ser216) (63F9) Rabbit mAb 对石蜡包埋的人结肠癌进行免疫组织化学分析,显示出胞浆定位。
    cdc25C Antibody Sampler Kit: Image 9 Expand Image
    使用 Phospho-cdc25C (Ser216) (63F9) Rabbit mAb 对石蜡包埋的人肺癌进行免疫组织化学分析。
    cdc25C Antibody Sampler Kit: Image 10 Expand Image
    使用 Phospho-cdc25C (Ser216) (63F9) Rabbit mAb 对未经处理(左)或经 λ 磷酸酶处理(右)的石蜡包埋的人肺癌进行免疫组织化学分析。
    cdc25C Antibody Sampler Kit: Image 11 Expand Image
    在对照肽(左)或 Phospho-cdc25C (Ser216) Blocking Peptide #1190(右)存在的情况下,使用 Phospho-cdc25C (Ser216) (63F9) Rabbit mAb 对石蜡包埋的人淋巴节点进行免疫组织化学分析。