PDGF Receptor α Antibody Sampler Kit

• 产品编号：CST-9360S      品牌：Cell Signaling Technology       原厂货号：9360S
• 产品分类：抗体 > 一抗 > 复合抗体试剂盒
• 应用分类：

其它组分：

描述：

PDGF Receptor α Antibody Sampler Kit中的每一种抗体能够检测其特异的靶分子在体内的表达水平，激活态抗体仅在特定位点磷酸化时识别靶蛋白。磷酸化PDGF Receptor α (Tyr849)/PDGF Receptor β (Tyr857) (C43E9) 兔单抗和 磷酸化PDGF Receptor α (Tyr1018) k抗体它激活的酪氨酸激酶蛋白发生交叉反应。PDGF Receptor α (D1E1E) XP® 兔单抗可能与高表达的PDGFRβ发生交叉反应。激活态的单克隆抗体通过用多肽免疫动物得到，该磷酸化多肽是根据以下位点附近的序列合成的：人 PDGFRα的Tyr754 和人PDGFRβ的Tyr857。总PDGFRα 单抗通过用多肽免疫动物得到，该多肽是根据人PDGFRα的羧基端序列合成的。激活态的多克隆抗体通过用多肽免疫动物得到，该磷酸化多肽是根据人 PDGFRα蛋白的Tyr1018位点附近的序列合成的。多克隆抗体经过protein A和亲和层析纯化。PDGF Receptor α Antibody Sampler Kit为检测PDGF receptor α protein (PDGFRα)和特定位点磷酸化的PDGFRα提供了经济有效地手段。本试剂盒的一抗和二抗足够四次免疫印迹检测实验使用。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

Each antibody in the PDGF Receptor α Antibody Sampler Kit detects endogenous levels of its target protein, with activation state antibodies recognizing target proteins only when phosphorylated at the indicated residues. Phospho-PDGF Receptor α (Tyr849)/PDGF Receptor β (Tyr857) (C43E9) Rabbit mAb and Phospho-PDGF Receptor α (Tyr1018) Antibody may cross-react with other activated tyrosine kinases; PDGF Receptor α (D1E1E) XP® Rabbit mAb may cross-react with PDGFRβ at overexpressed levels.

Source / Purification

Activation state monoclonal antibodies are produced by immunizing animals with synthetic phosphopeptides corresponding to residues surrounding Tyr754 of human PDGFRα and Tyr857 of human PDGFRβ. Total PDGFRα monoclonal antibody is produced by immunizing animals with a synthetic peptide corresponding to residues near the carboxy-terminal sequence of human PDGFRα. Activation state polyclonal antibodies are produced by immunizing animals with a synthetic phosphopeptide corresponding to residues surrounding Tyr1018 of human PDGFRα. Polyclonal antibodies are purified by protein A and peptide affinity chromatography.

Description

The PDGF Receptor α Antibody Sampler Kit provides an economical means of evaluating total PDGF receptor α protein (PDGFRα) levels as well as PDGFRα phosphorylated at specific sites. The kit contains enough primary and secondary antibody to perform four western blots with each antibody.

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: Supplied in 10 mM sodium HEPES (pH 7.5), 150
mM NaCl, 100 µg/ml BSA and 50% glycerol and less than
0.02% sodium azide. Store at –20°C. Do not aliquot the
antibodies.