| 货号 | 7316C |
| 描述 | The PathScan® Phospho-S6 Ribosomal Protein (Ser235/236) Chemiluminescent Sandwich ELISA Kit is a solid phase sandwich enzyme-linked immunosorbent assay (ELISA) that detects endogenous levels of phospho-S6 ribosomal protein (Ser235/236) with a chemiluminescent readout. Chemiluminescent ELISAs often have a wider dynamic range and higher sensitivity than conventional chromogenic detection. This chemiluminescent ELISA, which is offered in low volume microplates, shows increased signal and sensitivity while using smaller samples. A Phospho-S6 Ribosomal Protein (Ser235/236) Rabbit mAb has been coated on the microwells. After incubation with cell lysates, 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). Anti-mouse IgG, HRP-linked Antibody is then used to recognize the bound detection antibody. Chemiluminescent reagent is added for signal development. The magnitude of light emission, measured in relative light units (RLU), is proportional to the quantity of phospho-S6 ribosomal protein (Ser235/236). Antibodies in kit are custom formulations specific to kit. |
| 反应种属 | Human/Mouse |
| 应用 | ELISA |
| 目标/特异性 | The PathScan® Phospho-S6 Ribosomal Protein (Ser235/236) Chemiluminescent Sandwich ELISA Kit detects endogenous levels of phospho-S6 ribosomal protein phosphorylated on serines 235/236. This kit detects proteins from the indicated species, as determined through in-house testing, but may also detect homologous proteins from other species. |
| 供应商 | CST |
| 背景 | 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 (5TOP) 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). |
| 存放说明 | 4C |
| 参考文献 | Dufner, A. and Thomas, G. (1999) Exp Cell Res 253, 100-9. Peterson, R.T. and Schreiber, S.L. (1998) Curr Biol 8, R248-50. Jefferies, H.B. et al. (1997) EMBO J 16, 3693-704. Ferrari, S. et al. (1991) J Biol Chem 266, 22770-5. Flotow, H. and Thomas, G. (1992) J Biol Chem 267, 3074-8. |
Figure 1. Relationship between protein concentration from nonphospho cell lysate and PDGF-treated NIH/3T3 cell lysate and immediate light generation with chemiluminescent substrate is shown. Cells (80% confluence) lysed without the addition of phosphatase inhibitor to the lysis buffer (nonphospho lysate) or treated with PDGF (50 ng/ml) and lysed with Cell Lysis Buffer (10X) #9803 after incubation at 37ºC for 30 minutes. Graph inset corresponding to the shaded area shows high sensitivity and a linear response at the low protein concentration range. 图1. 非磷酸化细胞裂解物和PDGF处理的NIH/3T3细胞裂解物中的蛋白浓度与化学发光底物直接产生的光度值的关系如图所示。细胞(80% confluence)细胞(80% confluence)用没有加入磷酸化酶抑制剂的裂解液(nonphospho lysate) 裂解或 用hPDGF-BB #8912 (50 ng/ml, 37ºC时30分钟) 处理且加入细胞裂解液#9803进行裂解。相应的阴影面积插入的图片显示在低蛋白浓度区域具有较高的敏感度和一个线性的反应。 |
