| 货号 | 7082C |
| 描述 | The PathScan® Phospho-Insulin Receptor β (panTyr) Sandwich ELISA Kit from Cell Signaling Technology is a solid phase sandwich enzyme-linked immunosorbent assay (ELISA) that detects transfected levels of tyrosine-phosphorylated Insulin Receptor β protein. An Insulin Receptor β Rabbit mAb has been coated on the microwells. After incubation with cell lysates, Insulin Receptor β protein (phospho and nonphospho) is captured by the coated antibody. Following extensive washing, a phospho-tyrosine Mouse mAb is added to detect captured tyrosine-phosphorylated Insulin Receptor β protein. Anti-mouse IgG, HRP-linked antibody is then used to recognize the bound detection antibody. HRP substrate TMB is added to develop color. The magnitude of the absorbance for this developed color is proportional to the quantity of Insulin Receptor β protein phosphorylated on tyrosine. |
| 反应种属 | Human |
| 应用 | ELISA |
| 目标/特异性 | PathScan® Phospho-Insulin Receptor β (panTyr) Sandwich ELISA Kit #7082 detects transfected levels of human Insulin Receptor β protein phosphorylated at Tyrosine, as shown in Figure 1. The kit sensitivity is shown in Figure 2. 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 | Insulin receptor (INSR) is a membrane receptor tyrosine kinase. The receptor molecule consists of a disulfide linked heterodimer. The α subunit is a 135 kDa extracellular fragment, and the β subunit is a 95 kDa fragment containing an extracellular domain, a single transmembrane domain, and an intracellular tyrosine kinase domain (1). Insulin ligand binding to this receptor results in receptor autophosphorylation and tyrosine kinase activation. INSR catalyzes the tyrosine phosphorylation of molecules such as IRS, Gab1, Shc, and Cbl, which further activate the downstream MAPK, PI3K, and TC10 pathways. This eventually leads to increases in glucose uptake and metabolism as well as cell growth (2,3). INSR has peptide substrate specificity similar to other receptor tyrosine kinase members, preferring acidic residues at the -1 to -4 positions and large hydrophobic amino acids at positions +1 and +3 (4). |
| 存放说明 | 4C |
| 参考文献 | Yip, C.C. and Ottensmeyer, P. (2003) J. Biol. Chem. 278, 27329-27332. Saltiel, A.R. and Pessin, J.E. (2002) Trends Cell Biol. 12, 65-71. Zick, Y. (2001) Trends Cell Biol. 11, 437-441. Songyang, Z. and Cantley, L.C. (1995) TIBS 20, 470-475. |
Figure 1: Treatment of CHO-IR/IRS-1 cells with insulin stimulates tyrosine phosphorylation of Insulin Receptor β which is detected by the PathScan® Phospho-Insulin Receptor β (panTyr) Sandwich ELISA Kit #7082 (top, right). Similar levels of Insulin Receptor β protein from both nonphospho or phospho lysates are detected by PathScan® Total Insulin Receptor β Sandwich ELISA Kit #7069 (top, left). Absorbance at 450 nm is shown in the top figure while corresponding western blotsusing 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 the tranfected human insulin receptor and rat IRS-1. 图1，CHO-IR/IRS-1细胞经胰岛素处理后，胰岛素受体βTyr磷酸化水平升高，所用抗体为PathScan® Phospho-Insulin Receptor β (panTyr) Sandwich ELISA Kit #7082（右上）。用PathScan® Total Insulin Receptor β Sandwich ELISA Kit #7069（左上）检测磷酸化和非磷酸化的细胞裂解液中的胰岛素受体β水平没有差异。上图示450nm的吸光度，下图示对应的western blot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能够稳定表达人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 the absorbance at 450 nm is shown. After starvation, CHO-IR/IRS-1 cells (85% confluence) were treated with insulin (100 nM, 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。 |
