| 货号 | 7053S |
| 描述 | Cell Signaling Technologys PathScan® Phospho-p70 S6 Kinase (Thr389) Sandwich ELISA Antibody Pair is being offered as an economical alternative to our PathScan® Phospho-p70 S6 Kinase (Thr389) Sandwich ELISA Kit #7063. Capture and detection antibodies (100X stocks) and an HRP-conjugated secondary antibody (1000X stock) are supplied. Sufficient reagents are supplied for 4 x 96 well ELISAs. The p70 S6 kinase rabbit capture antibody is coated onto a 96 well microplate overnight in PBS. After blocking, cell lysates are added followed by a phospho-p70 S6 kinase (Thr389) mouse detection antibody and anti-mouse IgG, HRP-linked antibody. HRP substrate (TMB) is then added for color development. The magnitude of the absorbance for this developed color is proportional to the quantity of phospho-p70 S6 kinase (Thr389). |
| 反应种属 | Human/Mouse |
| 应用 | ELISA |
| 目标/特异性 | For Antibody Pair specificity and sensitivity, please refer to the corresponding PathScan® Sandwich ELISA Kit. Note: This antibody pair 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 | p70 S6 kinase is a mitogen activated Ser/Thr protein kinase that is required for cell growth and G1 cell cycle progression (1,2). p70 S6 kinase phosphorylates the S6 protein of the 40S ribosomal subunit and is involved in translational control of 5 oligopyrimidine tract mRNAs (1). A second isoform, p85 S6 kinase, is derived from the same gene and is identical to p70 S6 kinase except for 23 extra residues at the amino terminus, which encode a nuclear localizing signal (1). Both isoforms lie on a mitogen activated signaling pathway downstream of phosphoinositide-3 kinase (PI-3K) and the target of rapamycin, FRAP/mTOR, a pathway distinct from the Ras/MAP kinase cascade (1). The activity of p70 S6 kinase is controlled by multiple phosphorylation events located within the catalytic, linker and pseudosubstrate domains (1). Phosphorylation of Thr229 in the catalytic domain and Thr389 in the linker domain are most critical for kinase function (1). Phosphorylation of Thr389, however, most closely correlates with p70 kinase activity in vivo (3). Prior phosphorylation of Thr389 is required for the action of phosphoinositide 3-dependent protein kinase 1 (PDK1) on Thr229 (4,5). Phosphorylation of this site is stimulated by growth factors such as insulin, EGF and FGF, as well as by serum and some G-protein-coupled receptor ligands, and is blocked by wortmannin, LY294002 (PI-3K inhibitor) and rapamycin (FRAP/mTOR inhibitor) (1,6,7). Ser411, Thr421 and Ser424 lie within a Ser-Pro-rich region located in the pseudosubstrate region (1). Phosphorylation at these sites is thought to activate p70 S6 kinase via relief of pseudosubstrate suppression (1,2). Another LY294002 and rapamycin sensitive phosphorylation site, Ser371, is an in vitro substrate for mTOR and correlates well with the activity of a partially rapamycin resistant mutant p70 S6 kinase (8). |
| 存放说明 | 4C |
| 参考文献 | Pullen, N. and Thomas, G. (1997) FEBS Lett 410, 78-82. Dufner, A. and Thomas, G. (1999) Exp Cell Res 253, 100-9. Weng, Q.P. et al. (1998) J Biol Chem 273, 16621-9. Pullen, N. et al. (1998) Science 279, 707-10. Alessi, D.R. et al. (1998) Curr Biol 8, 69-81. Polakiewicz, R.D. et al. (1998) J Biol Chem 273, 23534-41. Fingar, D.C. et al. (2002) Genes Dev 16, 1472-87. Saitoh, M. et al. (2002) J Biol Chem 277, 20104-12. |
The relationship between the protein concentration of the lysate from untreated and IGF-1-treated MCF-7 cells and the absorbance at 450 nm using the PathScan® Phospho-p70 S6 Kinase (Thr389) Sandwich ELISA Antibody Pair #7053 is shown. MCF-7 cells were treated with 100 ng/ml hIGF-I #8917 for 20 minutes at 37ºC and then lysed. 采用IGF-1处理MCF7细胞和未处理组细胞提取物中蛋白浓度与450nm处吸光度值的关系图，所用试剂盒为PathScan ® Phospho-p70 S6 Kinase (Thr389) Sandwich ELISA Antibody Pair #7053 。MCF-7细胞采用100 ng/ml hIGF-I #8917在37度处理20分钟，然后裂解。 |
