PathScan? Phospho-Survivin (Thr34) Sandwich ELISA Kit

• 产品编号：CST-7193C      品牌：CST       原厂货号：7193C
• 产品分类：ELISA及其它免疫分析 > ELISA > ELISA检测试剂盒
• 应用分类：

描述：

CST的PathScan® Phospho-Survivin (Thr34) Sandwich ELISA Kit #7193产品能够检测内源性的Thr34处磷酸化的survivin蛋白，如图1所示。试剂盒灵敏度如图2所示。通过内部测试验证，本试剂盒能够检测指定物种的蛋白质，但也可以检测来自其他物种的同源蛋白。PathScan® Phospho-Survivin (Thr34) Sandwich ELISA Kit采用了固相夹心酶联免疫吸附法（ELISA）检测内源的Thr34磷酸化的 survivin 蛋白。微孔中包被了phospho-survivin (Thr34) rabbit antibody 。细胞裂解液孵育后，磷酸化 survivin 蛋白被包被的抗体捕获。充分洗涤后，添加survivin mouse antibody检测捕获的磷酸化survivin蛋白。再用Anti-mouse IgG, HRP-linked Antibody识别结合的抗体。添加HRP底物，TMB显色。测得的吸光值与Thr34磷酸化survivin蛋白的量成正比。Survivin是一个在胎儿发育和肿瘤细胞恶变过程中高表达的抗凋亡蛋白，大小为16 kDa（1）。Survivin结合并抑制caspase-3，通过抑制细胞凋亡调控和促进细胞分裂，从而来控制细胞周期的G2/M-期临界点（2,3）。这个调控过程需要Survivin在P34 CDC2激酶的作用下，在Thr34位发生磷酸化（4）。通过基因打靶实验和Survivin反义RNA实验，应用Thr34磷酸化缺陷的survivin突变体，已经证明了survivin能够抑制肿瘤的生长（5,6）。

1. Reed, J.C. and Reed, S.I. (1999) Nature Cell Biol. 1, 199-200.
2. Li, F. et al. (1998) Nature 396, 580-584.
3. Li, F. et al. (1999) Nat. Cell Biol. 1, 461-466.
4. O'Connor, D.S. et al. (2000) Proc. Natl. Acad. Sci. USA 97, 13103-13107.
5. Olie, R.A. et al. (2000) Cancer Res. 60, 2805-2809.
6. Grossman, D. et al. (2001) Proc. Natl. Acad. Sci. USA 98, 635-640.

原厂资料：

Specificity / Sensitivity

CST's PathScan® Phospho-Survivin (Thr34) Sandwich ELISA Kit #7193 detects endogenous levels of survivin protein phosphorylated at Thr34 as shown in Figure 1. Kit sensitivity is shown in Figure 2. This kit detects proteins from the indicated species, as determined through in-house testing, but may also detect homologous proteins from other species.

Description

The PathScan® Phospho-Survivin (Thr34) Sandwich ELISA Kit is a solid phase sandwich enzyme-linked immunosorbent assay (ELISA) that detects endogenous levels of survivin protein phosphorylated at Thr34. A phospho-survivin (Thr34) rabbit antibody has been coated onto the microwells. After incubation with cell lysates, phospho-survivin protein is captured by the coated antibody. Following extensive washing, a survivin mouse antibody is added to detect the captured phospho-survivin protein. Anti-mouse IgG, HRP-linked Antibody is then used to recognize the bound detection antibody. HRP substrate TMB is added to develop color. The magnitude of the absorbance for this developed color is proportional to the quantity of survivin phosphorylated at Thr34.

Background

Survivin is a 16 kDa anti-apoptotic protein highly expressed during fetal development and cancer cell malignancy (1). Survivin binds and inhibits caspase-3, controlling the checkpoint in the G2/M-phase of the cell cycle by inhibiting apoptosis and promoting cell division (2,3). This regulatory process requires the phosphorylation of survivin at Thr34 by p34 cdc2 kinase (4). Gene targeting using a Thr34 phosphorylation-defective survivin mutant, as well as antisense survivin, have been shown to inhibit tumor growth (5,6).

1. Reed, J.C. and Reed, S.I. (1999) Nature Cell Biol. 1, 199-200.
2. Li, F. et al. (1998) Nature 396, 580-584.
3. Li, F. et al. (1999) Nat. Cell Biol. 1, 461-466.
4. O'Connor, D.S. et al. (2000) Proc. Natl. Acad. Sci. USA 97, 13103-13107.
5. Olie, R.A. et al. (2000) Cancer Res. 60, 2805-2809.
6. Grossman, D. et al. (2001) Proc. Natl. Acad. Sci. USA 98, 635-640.

注意事项：

* 12 8-well modules -Each module is designed to break apart for 8 tests.
**Kit should be stored at 4°C with the exception of 10X Cell Lysis Buffer, which is stored at –20°C (packaged separately).