| 货号 | 7040C |
| 描述 | The PathScan® Total Axl Sandwich ELISA Kit is a solid phase sandwich enzyme-linked immunosorbent assay (ELISA) that detects endogenous levels of total Axl protein. An Axl mouse antibody has been coated on the microwells. After incubation with cell lysates, Axl protein (phospho and nonphospho) is captured by the coated antibody. Following extensive washing, an Axl rabbit antibody is added to detect captured Axl 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 the absorbance for this developed color is proportional to the quantity of total Axl protein. |
| 反应种属 | Human |
| 应用 | ELISA |
| 目标/特异性 | CSTs PathScan® Total Axl Sandwich ELISA Kit #7040 detects endogenous levels of total Axl protein in human cells, 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 | Axl, Sky and Mer are three members of a receptor tyrosine kinase (RTK) family that share a conserved intracellular tyrosine kinase domain and an extracellular domain similar to those seen in cell adhesion molecules. These RTKs bind the vitamin K-dependent protein growth-arrest-specific 6 (Gas6), which is structurally related to the protein S anticoagulation factor (1). Upon binding to its receptor, Gas6 activates phosphatidylinositol 3-kinase (PI3K) and its downstream targets Akt and S6K, as well as NF-κB (2,3). A large body of evidence supports a role for Gas6/Axl signaling in cell growth and survival in normal and cancer cells (4). |
| 存放说明 | 4C |
| 参考文献 | Crosier, K.E. and Crosier, P.S. (1997) Pathology 29, 131-5. Demarchi, F. et al. (2001) J Biol Chem 276, 31738-44. Lee, W.P. et al. (2002) Oncogene 21, 329-36. Bellosta, P. et al. (1997) Oncogene 15, 2387-97. |
Figure 1: Constitutive phosphorylation of Axl in NCI-H2347 cells lysed in the presence of phosphatase inhibitors* (phospho lysate) is detected by PathScan® Phospho-Axl (panTyr) Sandwich ELISA Kit #7042 (top, right). In contrast, a low level of phospho-Axl protein is detected in NCI-H2347 cells lysed in the absence of phosphatase inhibitors* (nonphospho lysate). Similar levels of total Axl protein from both nonphospho or phospho lysates are detected by PathScan® Total Axl Sandwich ELISA Kit #7040 (top, left). Absorbance at 450 nm is shown in the top figure while corresponding western blots using a phospho-Axl (Tyr814) rabbit antibody (right) or a total Axl (C44G1) Rabbit mAb #4566 (left) are shown in the bottom figure. *Phosphatase inhibitors include sodium pyrophosphate, β-glycerophosphate and Na3VO4.图1：在磷酸酶抑制剂*存在时（磷酸化的裂解液），用PathScan® Phospho-Axl (panTyr) Sandwich ELISA Kit #7042能够检测到NCI-H2347细胞中磷酸化Axl的组成型表达（上，右）。相比之下，没有磷酸酶抑制剂*存在时（非磷酸化的裂解液），仅能检测到低水平的磷酸化Axl的表达。用PathScan® Total Axl Sandwich ELISA Kit #7040则能在磷酸化和非磷酸化的裂解液中检测到近似水平的总Axl的表达（上，左）。上图显示450nm处的吸光值，下图显示用抗磷酸化Axl (Tyr814)的兔源单克隆抗体（右）或抗总Axl的兔源单克隆抗体(C44G1) #4566（左）进行免疫印迹检测的结果。*磷酸酶抑制剂中包含sodium pyrophosphate、β-glycerophosphate和 Na3VO4。 | |
Figure 2: The relationship between protein concentration of phospho or nonphospho lysates and the absorbance at 450 nm is shown. NCI-H2347 cells were cultured (85% confluence) and lysed with or without the addition of phosphatase inhibitor to the lysis buffer (phospho or nonphospho lysate).图2：磷酸化或非磷酸化的裂解中蛋白含量和450nm吸光值之间的关系。NCI-H2347细胞在85%满时收样，并用加入或未加磷酸酶抑制剂的裂解液进行处理（磷酸化或非磷酸化）。 |
