PathScan? Phospho-S6 Ribosomal Protein (Ser235/236) Sandwich ELISA Kit

• 产品编号：CST-7205C      品牌：CST       原厂货号：7205C
• 产品分类：ELISA及其它免疫分析 > ELISA > ELISA检测试剂盒
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描述：

PathScan® Phospho-S6 Ribosomal Protein (Ser235/236) Sandwich ELISA Kit仅检测在丝氨酸235/236位点磷酸化的内源性磷酸化S6核糖体蛋白。如图1所示，使用这个试剂盒检测后发现，PDGF处理的NIH/3T3细胞内的磷酸化S6核糖体蛋白(Ser235/236)蛋白显著的增加。然而，S6核糖体蛋白(磷酸化和非磷酸化)总蛋白水平仍然没有变化。NIH/3T3或293细胞在饥饿培养后加入20% FBS处理，也显示相似的结果（数据未显示）。通过内部实验确定，该试剂盒可检测指定物种，但是可能也会检测其它物种的同源性蛋白。该PathScan® Phospho-S6 Ribosomal Protein (Ser235/236) Sandwich ELISA Kit是一个固相夹心酶联免疫吸附法（ELISA），其能检测内源性磷酸化S6核糖体蛋白 (Ser235/236)。一个磷酸化S6核糖体蛋白 (Ser235/236)抗体已经被包被在微孔板上。在与细胞裂解液孵育后，仅磷酸化S6 核糖体蛋白被包被的抗体吸附。在充分清洗后，S6核糖体蛋白总蛋白的鼠单抗被加入来检测已被吸附的磷酸化S6核糖体蛋白 (Ser235/236)。然后，HRP-linked Anti-mouse Antibody被用于识别已结合的检测抗体。加入HRP底物TMB后显色。该色彩的吸光度值的高低与在磷酸化S6核糖体(Ser235/236)蛋白多少成正比。*试剂盒里的抗体是专门针对该试剂盒定制的。生长因子和有丝分裂原有效促进维持细胞生长和增殖的一个方面就是通过上调mRNA翻译(1,2)。生长因子和有丝分裂原能诱导p70 S6激酶的激活和随后S6核糖体蛋白的磷酸化。S6核糖体蛋白的磷酸化与mRNA转录本的翻译增加有关，而这个mRNA转录本是在5端非翻译区含有一个寡嘧啶区(2)。这些特殊的mRNA转录本 (5'TOP) 能够编码涉及细胞周期进程的蛋白质如翻译必须的核糖体蛋白和延伸因子(2,3)。重要的S6核糖体蛋白的磷酸化位点包含数个定位在S6蛋白一个小的羧基末端区域残基(Ser235, Ser236, Ser240, and Ser244) (4,5)。

1. Dufner, A. and Thomas, G. (1999) Exp Cell Res 253, 100-9.
2. Peterson, R.T. and Schreiber, S.L. (1998) Curr Biol 8, R248-50.
3. Jefferies, H.B. et al. (1997) EMBO J 16, 3693-704.
4. Ferrari, S. et al. (1991) J Biol Chem 266, 22770-5.
5. Flotow, H. and Thomas, G. (1992) J Biol Chem 267, 3074-8.

原厂资料：

Specificity / Sensitivity

The PathScan® Phospho-S6 Ribosomal Protein (Ser235/236) Sandwich ELISA Kit detects endogenous levels of phospho-S6 ribosomal protein phosphorylated on serines 235/236. As shown in Figure 1, a significant induction of phospho-S6 ribosomal protein (Ser235/236) in PDGF treated NIH/3T3 cells is detected by this kit. However, total S6 ribosomal protein (phospho and non-phospho) remains unchanged. NIH/3T3 or 293 cells treated with 20% FBS after starvation also show similar results (data not shown). 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-S6 Ribosomal Protein (Ser235/236) Sandwich ELISA Kit is a solid phase sandwich enzyme-linked immunosorbent assay (ELISA) that detects endogenous levels of phospho-S6 ribosomal protein (Ser235/236). A Phospho-S6 Ribosomal Protein (Ser235/236) Antibody has been coated onto the microwells. After incubation with cell lysates, only phospho-S6 ribosomal protein is captured by the coated antibody. Following extensive washing, a Total S6 Ribosomal Protein Mouse mAb is added to detect the captured phospho-S6 ribosomal protein (Ser235/236). HRP-linked Anti-mouse Antibody is then used to recognize the bound detection antibody. HRP substrate, TMB, is added to develop color. The magnitude of optical density for this developed color is proportional to the quantity of phospho-S6 ribosomal protein (Ser235/236). 
 * Antibodies in kit are custom formulations specific to kit.

Background

One way that growth factors and mitogens effectively promote sustained cell growth and proliferation is by upregulating mRNA translation (1,2). Growth factors and mitogens induce the activation of p70 S6 kinase and the subsequent phosphorylation of the S6 ribosomal protein. Phosphorylation of S6 ribosomal protein correlates with an increase in translation of mRNA transcripts that contain an oligopyrimidine tract in their 5' untranslated regions (2). These particular mRNA transcripts (5'TOP) encode proteins involved in cell cycle progression as well as ribosomal proteins and elongation factors necessary for translation (2,3). Important S6 ribosomal protein phosphorylation sites include several residues (Ser235, Ser236, Ser240, and Ser244) located within a small, carboxy-terminal region of the S6 protein (4,5).

1. Dufner, A. and Thomas, G. (1999) Exp Cell Res 253, 100-9.
2. Peterson, R.T. and Schreiber, S.L. (1998) Curr Biol 8, R248-50.
3. Jefferies, H.B. et al. (1997) EMBO J 16, 3693-704.
4. Ferrari, S. et al. (1991) J Biol Chem 266, 22770-5.
5. Flotow, H. and Thomas, G. (1992) J Biol Chem 267, 3074-8.

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).