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

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

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Supporting Data

REACTIVITY	H M R
SENSITIVITY	Endogenous
SOURCE	Rabbit

Application Key:

WB-Western Blotting

Species Cross-Reactivity Key:

H-Human
M-Mouse
R-Rat

Product Information

Product 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 an eIF4E antibody to control protein loading.

Protocol

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 independent of phosphorylation and is provided to control for protein loading.

Species Reactivity:

Human, Mouse, Rat

Source / Purification

Antibodies are produced by immunizing animals with synthetic peptides, and are purified by combinations of 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).

Database Links

UniProt ID: P06730 , P27361 , P31751 , Q9Y243 , P28482 , P31749 , Q06124 , P09619

Entrez-Gene Id: 1977 , 5595 , 208 , 10000 , 5594 , 207 , 5781 , 5159

Limited Uses

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For Research Use Only. Not For Use In Diagnostic Procedures.

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