| 货号 | 7302C |
| 描述 | CSTs PathScan® Phospho-IGF-I Receptor beta (Tyr1131) Sandwich ELISA Kit is a solid phase sandwich enzyme-linked immunosorbent assay (ELISA) that detects endogenous levels of IGF-I receptor beta protein when phosphorylated at Tyr1131. A Phospho-IGF-I Receptor beta (Tyr1131) Rabbit Antibody has been coated onto the microwells. After incubation with cell lysates, phospho-IGF-I Receptor beta is captured by the coated antibody. Following extensive washing, an IGF-I Receptor Mouse Antibody is added to detect the captured phospho-IGF-I receptor protein. Anti-mouse IgG, HRP-linked Antibody #7076 is then used to recognize the bound detection antibody. HRP substrate, TMB, is added to develop color. The magnitude of absorbance for this developed color is proportional to the quantity of IGF-I receptor protein phosphorylated at Tyr1131. Antibodies in kit are custom formulations specific to kit. |
| 反应种属 | Human |
| 应用 | ELISA |
| 目标/特异性 | CSTs PathScan® Phospho-IGF-I Receptor β (Tyr1131) Sandwich ELISA Kit #7302 detects endogenous levels of IGF-I Receptor β protein when phosphorylated at Tyr1131. A significant induction of phosphorylation of IGF-I receptor at Tyr1131 is detected in IGF-I- treated MCF-7 cells using PathScan® Phospho-IGF-I Receptor β (Tyr 1131) Sandwich ELISA Kit #7302 (Figure 1). The levels of total IGF-I receptor β (phospho and nonphospho) shown by Western analysis remain unchanged (Figure 1). 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 MCF-7 cells with IGF-I stimulates phosphorylation of IGF-I receptor at Tyr1131, detected by PathScan® Phospho-IGF-I Receptor β (Tyr1131) Sandwich ELISA Kit #7302, but does not affect the level of total IGF-I Receptor β protein detected by Western analysis. The absorbance readings at 450 nm are shown in the top figure, while the corresponding Western blots using Phospho-IGF-I Receptor β (Tyr1131)/Insulin Receptor β (Tyr1146) Antibody #3021 (right panel) or IGF-I Receptor β Antibody #3027 (left panel), are shown in the bottom figure. 图1，用IGF-I处理MCF-7细胞后，IGF-I受体β蛋白在1131位酪氨酸残基磷酸化水平明显提高，检测试剂盒为PathScan® Phospho-IGF-I Receptor β (Tyr1131) Sandwich ELISA Kit #7302，而Western检测所示总的IGF-I受体β蛋白（磷酸化和未磷酸化的）并没有改变。上图示450nm处的吸光度值，下图示相应的Western blot实验，所用抗体分别为Phospho-IGF-I Receptor β (Tyr1131)/Insulin Receptor β (Tyr1146) Antibody #3021（右）和IGF-I Receptor β Antibody #3027（左）。 | |
Figure 2. The relationship between the protein concentration of untreated and IGF-I-treated MCF-7 cell lysates and the absorbance at 450 nm is shown. Cells were serum starved overnight and then treated with 100 nm IGF-I for 5 min. at 37°C. MCF-7裂解液目标蛋白浓度与450nm处的吸光度值关系图，细胞未处理或用IGF-I处理，处理方法为：细胞血清饥饿过夜，然后用100nM IGF-I于37°C处理5min。 |
