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 Kit #7180

Kit Includes Quantity Applications Reactivity MW (kDa) Source
PathScan® PDGFR Activity Assay: Phospho-PDGFR, Phospho-SHP2, Phospho-Akt, and Phospho-p44/42 MAPK (Erk1/2) Multiplex Western Detection Cocktail II # 5304 250 microliters W H M R Rabbit
Treated and Untreated Control Cell Extracts 50 microliters
Anti-rabbit IgG, HRP-linked Antibody # 7074 25 microliters Goat
Anti-biotin, HRP-linked Antibody # 7075 50 microliters Goat
20X LumiGLO® Reagent and 20X Peroxide # 7003 2.5 milliliters
Biotinylated Protein Ladder Detection Pack # 7727 50 microliters

Applications Key:  W=Western Blotting
Reactivity Key:  H=Human  M=Mouse  R=Rat

Specificity / Sensitivity

Each phospho-antibody in this kit recognizes only the phosphorylated form of its specific target. The S6 Ribosomal Protein control antibody detects total levels of S6 to determine protein loading. All of the antibodies detect endogenous levels of the target proteins.

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: Multiplex Western Detection Kit to detect the phosphorylation of PDGFR, SHP2, Akt and p44/42 MAPK.

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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