PathScan? Phospho-PDGF Receptor α (Tyr849) Sandwich ELISA Antibody Pair

• 产品编号：CST-7317S      品牌：Cell Signaling Technology       原厂货号：7317S
• 产品分类：ELISA及其它免疫分析 > ELISA > ELISA检测抗体对
• 应用分类：

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描述：

抗体对的特异性和敏感性请参考相应的PathScan® Sandwich ELISA Kit。注意：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。CST公司的PathScan® Phospho-PDGF Receptor α (Tyr849) Sandwich ELISA Antibody Pair 是PathScan® Phospho-PDGF Receptor α (Tyr849) Sandwich ELISA Kit #7296的另一种经济型选择，主要提供捕获和检测抗体（100 X贮存液）以及HRP偶联二抗 (1000X贮存液），可供4 x 96孔ELISA检测使用。用PBS稀释后的磷酸化PDGFR α 兔捕获抗体包被96孔板，封闭后加入细胞裂解液孵育，然后加入PDGFR α 鼠检测抗体和HRP偶联二抗。最后加入HRP的底物TMB进行显色反应。按照相对光单位，发射光强度与磷酸化PDGFRα (Tyr849)蛋白的量成正比。*试剂盒中的抗体为试剂盒特别订制配方。血小板衍生生长因子(PDGF) 家族的蛋白以几种二硫键连接的二聚体形式存在 (PDGF AA, PDGF AB, PDGF BB, PDGF CC和PDGF DD)，它们以特定的方式与两种密切相关的酪氨酸激酶受体-PDGF receptor α (PDGFRα)和PDGF receptor β (PDGFRβ)-结合。PDGFRα 和PDGFRβ 的胞内激酶区有75% 到85% 的序列同源性，而激酶插入区和羧基端仅有很低的同源性(27% 到 28%) (1). PDGFRα 同型二聚体和除了含有PDGF D 之外的所有的PDGF亚型结合。PDGFRβ同型二聚体能结合PDGF BB 和DD亚型，还有PDGF AB 异二聚体。 异聚体 PDGF receptor α/β能结合 PDGF B, C, 和 D 同型二聚体，还有 PDGF AB 异二聚体 (2)。 PDGFRα 和 PDGFRβ 都能被PDGF激活和 EGFR形成异二聚体, (3)。各种细胞的受体总数、受体亚基组成都不相同，因此不同细胞对PDGF的反应也不同(4)。配体结合能诱发受体的二聚化和自磷酸化，继而结合并激活胞内含有SH2结构的信号传导分子， 如Grb2, Src, GAP, PI3K激酶, PLCγ和 Nck。激活的PDGFRs能引发一系列不同的信号通路 并影响细胞的生长、肌动蛋白重塑、迁移和分化 (5)。PDGFRβ激酶插入区的Tyr751位点结合PI3K激酶 (6)。根据PDGFRβ的Tyr751位点附近序列产生的磷酸化五肽(pTyr751-Val-Pro-Met-Leu)能够抑制PI3K激酶p85亚基的羧基端SH2区和PDGFRβ结合 (7)。 Tyr740对 PDGFRβ介导的 PI3 K激酶的激活也是必需的(8)。

1. Deuel, T.F. et al. (1988) Biofactors 1, 213-217.
2. Bergsten, E. et al. (2001) Nat. Cell Biol. 3, 512-516.
3. Betsholtz, C. et al. (2001) Bioessays 23, 494-507.
4. Coughlin, S.R. et al. (1988) Prog. Clin. Biol. Res. 266, 39-45.
5. Ostman, A. and Heldin, C.H. (2001) Adv. Cancer Res. 80, 1-38.
6. Panayotou, G. et al. (1992) EMBO J. 11, 4261-4272.
7. Ramalingam, K. et al. (1995) Bioorg. Med. Chem. 3, 1263-1272.
8. Kashishian, A. et al. (1992) EMBO J. 11, 1373-1382.

原厂资料：

Specificity / Sensitivity

For Antibody Pair specificity and sensitivity, please refer to the corresponding PathScan® Sandwich ELISA Kit. Note: This antibody pair detects proteins from the indicated species, as determined through in-house testing, but may also detect homologous proteins from other species.

Description

CST's PathScan® Phospho-PDGF Receptor α (Tyr849) Sandwich ELISA Antibody Pair is being offered as an economical alternative to our PathScan® Phospho-PDGF Receptor α (Tyr849) Sandwich ELISA Kit #7296. Capture and Detection Antibodies (100X stocks) and HRP-Conjugated Secondary Antibody (1000X stock) are supplied. Sufficient reagents are supplied for 4 x 96 well ELISAs. The Phospho-PDGFR α Rabbit Capture Antibody is coated in PBS overnight in a 96 well microplate. After blocking, cell lysates are added followed by PDGFR α Mouse Detection Antibody and HRP-conjugated Secondary Antibody. HRP substrate (TMB) is added for color development. The magnitude of the absorbance for this developed color is proportional to the quantity of phospho-PDGFRα (Tyr849) protein. 
 *Antibodies in this kit are custom formulations specific to the kit.

Background

The proteins of the platelet derived growth factor (PDGF) family exist as several disulphide-bonded, dimeric isoforms (PDGF AA, PDGF AB, PDGF BB, PDGF CC, and PDGF DD) that bind in a specific pattern to two closely related receptor tyrosine kinases, PDGF receptor α (PDGFRα) and PDGF receptor β (PDGFRβ). PDGFRα and PDGFRβ share 75% to 85% sequence homology between their two intracellular kinase domains while the kinase insert and carboxy-terminal tail regions display a lower level (27% to 28%) of homology (1). PDGFRα homodimers bind all PDGF isoforms except those containing PDGF D. PDGFRβ homodimers bind PDGF BB and DD isoforms, as well as the PDGF AB heterodimer. The heteromeric PDGF receptor α/β binds PDGF B, C, and D homodimers as well as the PDGF AB heterodimer (2). PDGFRα and PDGFRβ can each form heterodimers with EGFR, which is also activated by PDGF (3). Various cells differ in the total number of receptors present and in the receptor subunit composition, which may account for responsive differences among cell types to PDGF binding (4). Ligand binding induces receptor dimerization and autophosphorylation, followed by binding and activation of cytoplasmic SH2 domain-containing signal transduction molecules such as Grb2, Src, GAP, PI3 kinase, PLCγ, and Nck. A number of different signaling pathways are initiated by activated PDGF receptors and lead to control of cell growth, actin reorganization, migration, and differentiation (5). Tyr751 in the kinase-insert region of PDGFRβ is the docking site for PI3 kinase (6). Phosphorylated pentapeptides derived from Tyr751 of PDGFRβ (pTyr751-Val-Pro-Met-Leu) inhibit the association of the carboxy-terminal SH2 domain of the p85 subunit of PI3 kinase with PDGFRβ (7). Tyr740 is also required for PDGFRβ-mediated PI3 kinase activation (8).

1. Deuel, T.F. et al. (1988) Biofactors 1, 213-217.
2. Bergsten, E. et al. (2001) Nat. Cell Biol. 3, 512-516.
3. Betsholtz, C. et al. (2001) Bioessays 23, 494-507.
4. Coughlin, S.R. et al. (1988) Prog. Clin. Biol. Res. 266, 39-45.
5. Ostman, A. and Heldin, C.H. (2001) Adv. Cancer Res. 80, 1-38.
6. Panayotou, G. et al. (1992) EMBO J. 11, 4261-4272.
7. Ramalingam, K. et al. (1995) Bioorg. Med. Chem. 3, 1263-1272.
8. Kashishian, A. et al. (1992) EMBO J. 11, 1373-1382.

注意事项：

Storage: Capture and Detection Antibodies are stored at
4ºC.HRP-Conjugated Secondary Antibody is stored at -20ºC.