PathScan? Bcr/Abl Activity Assay: Phospho-c-Abl, Phospho-Stat5 and Phospho-CrkL Multiplex Western De

• 产品编号：CST-5300S      品牌：CST       原厂货号：5300S
• 产品分类：抗体 > 一抗 > 磷酸化抗体
• 应用分类：

描述：

cocktail中的每个磷酸化抗体都能识别内源性特异靶点磷酸化的蛋白水平。eIF4E Antibody都能检测它的内源性靶蛋白水平，并且不依赖于磷酸化，也能用于对照。该多克隆抗体通过使用合成肽段免疫动物而获得。该抗体经蛋白A和肽亲和层析纯化。PathScan® Multiplex Western Detection Cocktail为分析一种胞膜上各种蛋白的抑制提供了独特的方法，不用去除免疫印迹膜上的封闭液和抗体，然后再次重新标记。该方法为使用者节约了时间，且调高了准确度，减少了试剂的浪费。Bcr/Abl Activity Assay让使用者能即时检测应答STI-571后c-Abl, Stat5，CrkL蛋白的磷酸化抑制。Cocktail也包括用于控制蛋白负载的elF4E antibody。STI-571 (也被称为Imatinib mesylate and Gleevec™)是一种酪氨酸激酶（TK）抑制子，它是 Bcr-Abl, PDGF受体和c-Kit TKs的相对特异ATP结合位点拮抗物(1-3)。CML临床试验效果很好，STI-571已经成为靶点癌症治疗的典范(4-6)。通过磷酸化酪氨酸途径的信号转导目前已经有许多人在进行研究，酪氨酸的磷酸化与许多细胞生长和分化途径有关(7-9)。因为观察到的CML造血功能依赖于未受损的Bcr-Abl酪氨酸激酶活性，目前对于Bcr-Abl底物的鉴定和脊髓扩张的可能机制已经做了许多研究。许多研究者描述了CML blasts和Bcr-Abl-表达细胞系中主要酪氨酸磷酸化的蛋白底物，这些细胞系包括SHIP, c-cbl, Dok, SHC，CrkL (10-15)。除此之外，涉及PI3 kinase, Ras, Myc和Stat5的关键信号转导通路也以Bcr-Abl激酶依赖型方式被激活(16)。

1. Buchdunger, E. et al. (1996) Cancer Res 56, 100-4.
2. Heinrich, M.C. et al. (2000) Blood 96, 925-32.
3. Druker, B.J. et al. (1996) Nat Med 2, 561-6.
4. Mauro, M.J. and Druker, B.J. (2001) Curr Oncol Rep 3, 223-7.
5. Druker, B.J. et al. (2001) N Engl J Med 344, 1031-7.
6. Druker, B.J. et al. (2001) N Engl J Med 344, 1038-42.
7. Blume-Jensen, P. and Hunter, T. (2001) Nature 411, 355-65.
8. Ullrich, A. and Schlessinger, J. (1990) Cell 61, 203-12.
9. Cantley, L.C. et al. (1991) Cell 64, 281-302.
10. ten Hoeve, J. et al. (1994) Blood 84, 1731-6.
11. Matsuguchi, T. et al. (1994) J Biol Chem 269, 5016-21.
12. Carpino, N. et al. (1997) Cell 88, 197-204.
13. Sattler, M. et al. (1997) Oncogene 15, 2379-84.
14. Di Cristofano, A. et al. (1998) J Biol Chem 273, 4827-30.
15. Wisniewski, D. et al. (1999) Blood 93, 2707-20.
16. Kabarowski, J.H. and Witte, O.N. (2000) Stem Cells 18, 399-408.

原厂资料：

Specificity / Sensitivity

Each phospho-antibody in this cocktail recognizes endogenous levels of only the phosphorylated form of its specific target. The eIF4E Antibody detects endogenous levels of its target protein independent of phosphorylation and is provided to control for protein loading.

Source / Purification

Polyclonal antibodies are produced by immunizing animals with synthetic peptides. Antibodies are purified by protein A and peptide affinity chromatography.

Description

The PathScan® Multiplex Western Detection Cocktail offers a unique method to assay the inhibition of multiple proteins on one membrane without stripping and reprobing. This method saves the user valuable time while increasing accuracy and minimizing reagent waste. The Bcr/Abl Activity Assay allows the user to simultaneously detect the inhibition of phosphorylation of c-Abl, Stat5 and CrkL proteins in response to STI-571. The cocktail also includes elF4E antibody to control protein loading.

Background

STI-571 (also known as Imatinib mesylate and Gleevec™) is a tyrosine kinase (TK) inhibitor that is a relatively specific ATP-binding site antagonist of Bcr-Abl, PDGF receptor and c-Kit TKs (1-3). Results are encouraging in CML clinical trials, and STI-571 has become a paradigm for targeted cancer therapeutics (4-6). Signal transduction through phospho-tyrosine pathways has been studied extensively, and tyrosine phosphorylation has been linked to multiple cell growth and differentiation pathways (7-9). Because the observed leukemic state of CML is dependent on the intact Bcr-Abl tyrosine kinase activity, extensive work has been done to identify substrates of Bcr-Abl and thus possible mechanisms leading to a myeloid expansion. Many groups have characterized prominent tyrosine-phosphorylated protein substrates in both CML blasts and Bcr-Abl-expressing cell lines, including SHIP, c-cbl, Dok, SHC and CrkL (10-15). In addition, key signal transduction pathways involving PI3 kinase, Ras, Myc and Stat5 are also activated in a Bcr-Abl kinase-dependent manner (16).

1. Buchdunger, E. et al. (1996) Cancer Res 56, 100-4.
2. Heinrich, M.C. et al. (2000) Blood 96, 925-32.
3. Druker, B.J. et al. (1996) Nat Med 2, 561-6.
4. Mauro, M.J. and Druker, B.J. (2001) Curr Oncol Rep 3, 223-7.
5. Druker, B.J. et al. (2001) N Engl J Med 344, 1031-7.
6. Druker, B.J. et al. (2001) N Engl J Med 344, 1038-42.
7. Blume-Jensen, P. and Hunter, T. (2001) Nature 411, 355-65.
8. Ullrich, A. and Schlessinger, J. (1990) Cell 61, 203-12.
9. Cantley, L.C. et al. (1991) Cell 64, 281-302.
10. ten Hoeve, J. et al. (1994) Blood 84, 1731-6.
11. Matsuguchi, T. et al. (1994) J Biol Chem 269, 5016-21.
12. Carpino, N. et al. (1997) Cell 88, 197-204.
13. Sattler, M. et al. (1997) Oncogene 15, 2379-84.
14. Di Cristofano, A. et al. (1998) J Biol Chem 273, 4827-30.
15. Wisniewski, D. et al. (1999) Blood 93, 2707-20.
16. Kabarowski, J.H. and Witte, O.N. (2000) Stem Cells 18, 399-408.

注意事项：

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