| 货号 | 7258C |
| 描述 | CSTs PathScan® Phospho-Insulin Receptor β (Tyr1150/1151) Sandwich ELISA Kit is a solid phase sandwich enzyme-linked immunosorbent assay (ELISA) that detects transfected phospho-insulin receptor (Tyr1150/1151) protein. An Insulin Receptor β Mouse mAb has been coated onto the microwells. After incubation with cell lysates, both phospho- and nonphospho-insulin receptor proteins are captured by the coated antibody. Following extensive washing, Phospho-IGF-I Receptor β (Tyr1135/1136)/Insulin Receptor β (Tyr1150/1151) (19H7) Rabbit mAb is added to detect the captured phospho-insulin receptor (Tyr1150/1151) protein. Anti-rabbit IgG, HRP-linked 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-insulin receptor β (Tyr1150/1151) protein. * Antibodies in kit are custom formulations specific to kit. |
| 反应种属 | Human/Mouse |
| 应用 | ELISA |
| 目标/特异性 | CSTs PathScan® Phospho-Insulin Receptor β (Tyr1150/1151) Sandwich ELISA Kit #7258 detects phospho-insulin receptor (Tyr1150/1151) protein. As shown in Figure 1, using Phospho-Insulin Receptor β (Tyr1150/1151) Sandwich ELISA Kit #7258, a significant induction of phospho-insulin receptor (Tyr1150/1151) is detected in CHO-IR/IRS-1 cells treated with insulin. The levels of total insulin receptor β (phospho and nonphospho) shown by Western analysis remain unchanged. Endogenous phospho-insulin receptor β (Tyr1150/1151) in either NIH/3T3 or HepG2 cells treated with insulin is also detected by Phospho-Insulin Receptor β (Tyr1150/1151) Sandwich ELISA Kit #7258. 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 | Type I insulin-like growth factor receptor (IGF-IR) is a transmembrane receptor tyrosine kinase that is widely expressed in many cell lines and cell types within fetal and postnatal tissues (1-3). Receptor autophosphorylation follows binding of the IGF-I and IGF-II ligands. Three tyrosine residues within the kinase domain (Tyr1131, Tyr1135, and Tyr1136) are the earliest major autophosphorylation sites (4). Phosphorylation of these three tyrosine residues is necessary for kinase activation (5,6). Insulin receptors (IRs) share significant structural and functional similarity with IGF-I receptors, including the presence of an equivalent tyrosine cluster (Tyr1146/1150/1151) within the kinase domain activation loop. Tyrosine autophosphorylation of IRs is one of the earliest cellular responses to insulin stimulation (7). Autophosphorylation begins with phosphorylation of Tyr1146 and either Tyr1150 or Tyr1151, while full kinase activation requires triple tyrosine phosphorylation (8). |
| 存放说明 | 4C |
| 参考文献 | Adams, T.E. et al. (2000) Cell Mol Life Sci 57, 1050-93. Baserga, R. (2000) Oncogene 19, 5574-81. Scheidegger, K.J. et al. (2000) J Biol Chem 275, 38921-8. Hernández-Sánchez, C. et al. (1995) J Biol Chem 270, 29176-81. Lopaczynski, W. et al. (2000) Biochem Biophys Res Commun 279, 955-60. Baserga, R. (1999) Exp Cell Res 253, 1-6. White, M.F. et al. (1985) J Biol Chem 260, 9470-8. White, M.F. et al. (1988) J Biol Chem 263, 2969-80. |
Figure 1: Treatment of CHO-IR/IRS-1 cells with insulin stimulates phosphorylation of insulin receptor at Tyr1150/1151, detected by PathScan® Phospho-Insulin Receptor β (Tyr1150/1151) Sandwich ELISA Kit #7258, but the level of total insulin receptor shown by Western analysis remains unchanged. OD 450 nm eadings are shown in the top figure, while the corresponding Western blots using Phospho-IGF-I Receptor β (Tyr1135/1136)/Insulin Receptor β (Tyr1150/1151) (19H7) Rabbit mAb #3024 (right panel) or Insulin Receptor β Antibody #3025 (left panel), are shown in the bottom figure. CHO-IR/IRS-1 cells stably overexpress human insulin receptor and rat IRS-1. 1图所示为CHO-IR/IRS-1细胞经胰岛素处理后，Tyr1150/1151磷酸化的胰岛素受体水平明显提高，所用抗体为Phospho-Insulin Receptor β (Tyr1150/1151) Sandwich ELISA Kit #7258。但是Western检测所示胰岛素受体β（磷酸化和未磷酸化的）总水平没有变化。上图示450nm处的吸光度值，下图为对应的Western实验，所用抗体为Phospho-IGF-I Receptor β (Tyr1135/1136)/Insulin Receptor β (Tyr1150/1151) (19H7) Rabbit mAb兔单抗 #3024（右）和Insulin Receptor β Antibody #3025（左）。CHO-IR/IRS-1稳转人的胰岛素受体和大鼠胰岛素受体底物1。 | |
Figure 2: The relationship between protein concentration of lysates from untreated and insulin-treated CHO-IR/IRS-1 cells and kit assay optical density readings is shown. After starvation, CHO-IR/IRS-1 cells (85% confluence) were treated with insulin (100 nM) for 2 min at 37°C and then lysed. CHO-IR/IRS-1细胞裂解液蛋白浓度与450nm处的吸光度关系图，细胞未处理或经胰岛素处理。处理方法如下：细胞（85%）饥饿后，用100 nM胰岛素于37度下处理2分钟，然后进行裂解。 |
