Cell Signaling Technology

Product Pathways - MAPK Signaling

PathScan® PDGFR Activity Assay: Phospho-PDGFR, Phospho-SHP2, Phospho-Akt, and Phospho-p44/42 MAPK (Erk1/2) Multiplex Western Detection Cocktail II #5304

Applications Reactivity Source
W H M R Rabbit

Applications Key:  W=Western Blotting
Reactivity Key:  H=Human  M=Mouse  R=Rat
Species cross-reactivity is determined by Western blot.

This Cocktail Contains The Following Antibodies:

Western Blotting

Western Blotting

Western blot analysis of extracts from serum-starved NIH/3T3 cells untreated or PDGF-treated (#9909, 100 ng/ml for 10 minutes), using PathScan PDGF Receptor Activity Assay cocktail to detect phosphorylation of PDGFR, SHP2, Akt and p44/42 MAPK.

Description

The PathScan® Multiplex Western Detection Cocktail offers a unique method to assay the activation 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 PDGFR Tyrosine Kinase Activity Assay allows the user to simultaneously detect the phosphorylation of PDGF receptor, SHP2, Akt and p44/42 MAPK proteins in response to PDGF. The kit also includes S6 Ribosomal Protein 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 S6 Ribosomal Protein Antibody detects endogenous levels of its target 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

Platelet-derived growth factor (PDGF) is a dimeric molecule that exists as homodimers or heterodimers of related polypeptide chains (A and B). Two types of PDGF receptors have been identified. The PDGF alpha-receptor binds all three isoforms with high affinity, whereas the beta-receptor binds only PDGF-BB with high affinity, PDGF-AB with low affinity and does not appear to bind PDGF-AA (1). PDGF exerts its stimulatory effects on cells by binding to these two related protein tyrosine kinase receptors. Ligand binding induces receptor dimerization and autophosphorylation, allowing binding and activation of cytoplasmic SH2-domain-containing signal transduction molecules. Thereby, a number of different signaling pathways are initiated, leading to cell growth, actin reorganization, migration and differentiation (2-4). In clinical studies, PDGF expression has been shown in a number of different solid tumors, from glioblastomas to prostate carcinomas. In these various tumor types, the biologic role of PDGF signaling can vary from autocrine stimulation of cancer cell growth to more subtle paracrine interactions involving adjacent stroma and even angiogenesis. Targeting PDGF signaling becomes an effective way for tumor treatment (5).

  1. Westermark, B. et al. (1990) Ciba Found. Symp. 150, 6-22.
  2. Heldin, C.H. (1997) FEBS Lett. 410, 17-21.
  3. Bornfeldt, K. E. et al. (1995) Ann. N. Y. Acad. Sci. 766, 416-430.
  4. Renhowe, P.A. (2002) Curr. Opin. Drug Discov. Devel. 5, 214-224.
  5. George, D. (2001) Semin. Oncol. 28, 27-33.

Application References

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Companion Products

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.

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