Product Pathways - PathScan Multiplex WB Cocktails
PathScan® Bcr/Abl Activity Assay: Phospho-c-Abl, Phospho-Stat5 and Phospho-CrkL Multiplex Western Detection Cocktail #5300
Reactivity Key: H=Human
Species cross-reactivity is determined by western blot. Species enclosed in parentheses are predicted to react based on 100% sequence homology.
This Cocktail Contains The Following Antibodies:
- Phospho-c-Abl (Tyr245) (73E5) Rabbit mAb #2868
- Phospho-Stat5 (Tyr694) (D47E7) XP® Rabbit mAb #4322
- Phospho-CrkL (Tyr207) Antibody #3181
- Rab11 (D4F5) XP® Rabbit mAb #5589
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 Rab11 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 Rab11 Antibody detects endogenous levels of its target protein independent of phosphorylation and is provided to control for protein loading.
Source / Purification
Antibodies are produced by immunizing animals with synthetic peptides. Polyclonal antibodies are purfied by protein A and peptide affinity chromatography.
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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- 2868 Phospho-c-Abl (Tyr245) (73E5) Rabbit mAb
- 4322 Phospho-Stat5 (Tyr694) (D47E7) XP® Rabbit mAb
- 3181 Phospho-CrkL (Tyr207) Antibody
- 5589 Rab11 (D4F5) XP® Rabbit mAb
- 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
Rabbit Monoclonals Produced Using Technology from Epitomics, Inc. Under U.S. patent No. 5,675,063.
For Research Use Only. Not For Use In Diagnostic Procedures.