Product Pathways - PathScan Multiplex WB Cocktails
PathScan® Bcr/Abl Activity Assay: Phospho-c-Abl, Phospho-Stat5 and Phospho-CrkL Multiplex Western Detection Cocktail II #5300
| Applications | Reactivity | Source |
|---|---|---|
| W | H | Rabbit |
Applications Key:
W=Western Blotting
Reactivity Key:
H=Human
Species cross-reactivity is determined by Western blot.
This Cocktail Contains The Following Antibodies:
- Phospho-c-Abl (Tyr245) Antibody #2861
- Phospho-Stat5 (Tyr694) Antibody #9351
- Phospho-CrkL (Tyr207) Antibody #3181
- eIF4E Antibody #9742
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.
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 rabbits with synthetic peptides. Antibodies are purified by protein A and peptide affinity chromatography.
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).
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- Druker, B.J. et al. (2001) N Engl J Med 344, 1031-7.
- Druker, B.J. et al. (2001) N Engl J Med 344, 1038-42.
- Blume-Jensen, P. and Hunter, T. (2001) Nature 411, 355-65.
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Application References
Have you published research involving the use of our products? If so we'd love to hear about it. Please let us know!
Companion Products
- 9351 Phospho-Stat5 (Tyr694) Antibody
- 3181 Phospho-CrkL (Tyr207) Antibody
- 2861 Phospho-c-Abl (Tyr245) Antibody
- 7074 Anti-rabbit IgG, HRP-linked Antibody
- 7071 Phototope®-HRP Western Blot Detection System, Anti-rabbit IgG, HRP-linked Antibody
- 7727 Biotinylated Protein Ladder Detection Pack
- 7720 Prestained Protein Marker, Broad Range (Premixed Format)
- 7003 20X LumiGLO® Reagent and 20X Peroxide
- 7130 PathScan® Bcr/Abl Activity Assay: Phospho-c-Abl, Phospho-Stat5 and Phospho-CrkL Multiplex Western Detection Kit
This product is for in vitro research use only and is not intended for use in humans or animals. This product is not intended for use as therapeutic or in diagnostic procedures.