PathScan? Phospho-Axl (panTyr) Sandwich ELISA Kit

• 产品编号：CST-7042C      品牌：CST       原厂货号：7042C
• 产品分类：ELISA及其它免疫分析 > ELISA > ELISA检测试剂盒
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描述：

CST公司的磷酸化Axl (panTyr) Sandwich ELISA Kit #7042用于检测人细胞中内源性酪氨酸被磷酸化的Axl的表达（如图1）。图2显示试剂盒的灵敏度。内部检测显示，本试剂盒能检测列举出的物种蛋白，但不排除用于其它物种同源蛋白检测的可能。PathScan® 磷酸化Axl (panTyr) Sandwich ELISA Kit利用固相双抗夹心酶联免疫检测法对内源性酪氨酸被磷酸化的Axl蛋白进行检测。微孔中已经包被了抗Axl的小鼠源抗体，与细胞裂解液共孵育后，磷酸化的和未磷酸化的Axl均能被包被抗体捕获。多次洗板之后，加入抗酪氨酸被磷酸化的Axl的兔源抗体对被捕获的酪氨酸被磷酸化的Axl蛋白进行检测，然后加入HRP标记的抗兔的IgG对检测抗体进行识别，最后加入HRP的底物TMB进行显色反应。按照相对光单位，发射光强度与酪氨酸位点被磷酸化的Axl蛋白量成正比。Axl、 Sky和 Mer是 受体酪氨酸激酶 (RTK) 家族的三个成员，它们有共同的胞内酪氨酸激酶区域和与胞外粘附分子相似的胞外区。这些受体酪氨酸激酶与维生素K依赖性的生长停滞特异性6蛋白（Gas6）结合，Gas6蛋白与S抗凝血因子蛋白结构上相关(1)。与受体结合后，Gas6能够激活PI3K激酶和它的下游靶分子Akt、 S6K和 NF-κB (2,3)。大量证据表明Gas6/Axl信号通路与正常细胞和癌细胞的生长和存活相关(4)。

1. Crosier, K.E. and Crosier, P.S. (1997) Pathology 29, 131-135.
2. Demarchi, F. et al. (2001) J. Biol. Chem. 276, 31738-31744.
3. Lee, W. P. et al. (2002) Oncogene 21, 329-336.
4. Bellosta, P. et al. (1997) Oncogene 15, 2387-2397.

原厂资料：

Specificity / Sensitivity

CST's PathScan® Phospho-Axl (panTyr) Sandwich ELISA Kit #7042 detects endogenous levels of tyrosine-phosphorylated 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.

Description

The PathScan® Phospho-Axl (panTyr) Sandwich ELISA Kit is a solid phase sandwich enzyme-linked immunosorbent assay (ELISA) that detects endogenous levels of tyrosine-phosphorylated 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, a phospho-tyrosine rabbit antibody is added to detect captured tyrosine-phosphorylated 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 Axl protein phosphorylated on tyrosine residues.

Background

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).

1. Crosier, K.E. and Crosier, P.S. (1997) Pathology 29, 131-135.
2. Demarchi, F. et al. (2001) J. Biol. Chem. 276, 31738-31744.
3. Lee, W. P. et al. (2002) Oncogene 21, 329-336.
4. Bellosta, P. et al. (1997) Oncogene 15, 2387-2397.

注意事项：

* 12 8-well modules -Each module is designed to break apart for 8 tests.
**Kit should be stored at 4°C with the exception of 10X Cell Lysis Buffer, which is stored at –20°C (packaged separately).