PathScan? Total Ret Sandwich ELISA Kit

• 产品编号：CST-7032C      品牌：CST       原厂货号：7032C
• 产品分类：ELISA及其它免疫分析 > ELISA > ELISA检测试剂盒
• 应用分类：

其它组分：

描述：

PathScan® Total Ret Sandwich ELISA Kit #7032用于检测人细胞中内源总Ret蛋白的表达（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® Total Ret Sandwich ELISA Kit利用固相双抗夹心酶联免疫检测法(ELISA)对内源性总Ret的表达进行检测。微孔中已经包被了Ret兔单抗，与细胞裂解液共孵育后，磷酸化的和未磷酸化的Ret蛋白均能被包被抗体捕获。多次洗板之后，加入抗体Ret鼠单抗对被捕获的Ret蛋白进行检测，然后加入抗鼠IgG, HRP偶联抗体对结合上的检测抗体进行识别，最后加入HRP的底物TMB进行显色反应。按照相对光单位，发射光强度与总Ret的表达量成正比。Ret原癌基因（c-Ret）是一个多功能的酪氨酸激酶受体复合物，它和其它膜结合的GDNF家族的配体结合型受体共同发挥作用（1）。与Ret受体结合的配体包括神经胶质细胞来源的神经营养因子（GDNF）和它的同类物neurturin、 persephin 和 artemin (2-4)。在相应的Ret 基因的改变会引起乳突状甲状腺癌、多发性内分泌肿瘤（2A和2B型），家族性髓样甲状腺癌和先天性巨结肠病(1,3)。位于Ret激酶区的Tyr905位点对于Ret的催化功能和生物学功能十分重要。替换Tyr905的Phe能显著抑制Ret的自磷酸化活性（5）。

1. Airaksinen, M.S. et al. (1999) Mol Cell Neurosci 13, 313-25.
2. Takahashi, M. et al. (1989) Oncogene 4, 805-6.
3. Manié, S. et al. (2001) Trends Genet 17, 580-9.
4. Tallini, G. and Asa, S.L. (2001) Adv Anat Pathol 8, 345-54.
5. Iwashita, T. et al. (1999) Oncogene 18, 3919-22.

原厂资料：

Specificity / Sensitivity

PathScan® Total Ret Sandwich ELISA Kit #7032 detects endogenous levels of total Ret 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® Total Ret Sandwich ELISA Kit is a solid phase sandwich enzyme-linked immunosorbent assay (ELISA) that detects endogenous levels of total Ret protein. A Ret rabbit mAb has been coated on the microwells. After incubation with cell lysates, Ret protein (phospho and nonphospho) is captured by the coated antibody. Following extensive washing, a Ret mouse mAb is added to detect captured Ret 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 total Ret protein.

Background

The Ret proto-oncogene (c-Ret) is a receptor tyrosine kinase that functions as a multicompetent receptor complex in conjunction with other membrane-bound ligand-binding GDNF family receptors (1). Ligands that bind the Ret receptor include the glial cell line-derived neurotropic factor (GDNF) and its congeners neurturin, persephin and artemin (2-4). Alterations in the corresponding Ret gene are associated with diseases including papillary thyroid carcinoma, multiple endocrine neoplasia (type 2A and 2B), familial medullary thyroid carcinoma and a congenital developmental disorder known as Hirschsprung’s disease (1,3). The Tyr905 residue located in the Ret kinase domain plays a crucial role in Ret catalytic and biological activity. Substitution of Phe for Tyr905 dramatically inhibits Ret autophosphorylation activity (5).

1. Airaksinen, M.S. et al. (1999) Mol Cell Neurosci 13, 313-25.
2. Takahashi, M. et al. (1989) Oncogene 4, 805-6.
3. Manié, S. et al. (2001) Trends Genet 17, 580-9.
4. Tallini, G. and Asa, S.L. (2001) Adv Anat Pathol 8, 345-54.
5. Iwashita, T. et al. (1999) Oncogene 18, 3919-22.

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).