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Assay Type
Solid Phase Sandwich ELISA
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Format
96-well strip plate
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Sample Type & Volume Required
Cell lysates (100 µL)
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Sufficient Materials
Kits available for two, five, or fifteen 96-well plates*
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Specificity
Please see the
* Provided that the recommended microplates, buffers, diluents, substrates and solutions are used, and the assay is run as summarized in the Assay Procedure provided.
This DuoSet IC ELISA contains the basic components required for the development of sandwich ELISAs to measure tyrosine-phosphorylated in cell lysates. An immobilized capture antibody specific for binds both phosphorylated and unphosphorylated . After washing away unbound material, a biotinylated detection antibody is used to detect only phosphorylated receptor, utilizing a standard HRP format.
Product Features
- Optimized capture and detection antibody pairings and recommended concentrations save lengthy development time
- Development protocols are provided to guide further assay optimization
- Assay can be customized to your specific needs
- Available in 2, 5, and 15-(96-well) plate pack sizes
- Economical alternative to Western blot
Kit Content
- Capture Antibody
- Conjugated Detection Antibody
- Calibrated Immunoassay Standard or Control
Other Reagents Required
PBS: (Catalog # ), or 137 mM NaCl, 2.7 mM KCl, 8.1 mM Na2HPO4, 1.5 mM KH2O4, pH 7.2 - 7.4, 0.2 µm filtered
Wash Buffer: (Catalog # ), or equivalent
Lysis Buffer*
IC Diluent*
Blocking Buffer*
Substrate Solution: 1:1 mixture of Color Reagent A (H2O2) and Color Reagent B (Tetramethylbenzidine) (Catalog # )
Stop Solution: 2 N H2SO4 (Catalog # )
Microplates: R&D Systems (Catalog # ), or equivalent
Plate Sealers: ELISA Plate Sealers (Catalog # ), or equivalent
*For the Lysis Buffer, IC Diluent, and Blocking BUffer recommended for a specific DuoSet ELISA Development Kit, please see the product
Preparation and Storage
Background: PDGF R beta
PDGF isoforms exert their cellular effects by binding to the structurally similar receptor tyrosine kinases (RTK), PDGF R alpha (CD140a) and PDGF R beta (CD140b). PDGF-AA, PDGF-BB, PDGF-CC, PDGF-DD and PDGF-AB isofoms bind the two distinct cell surface PDGF receptors with different affinities. Both PDGF R alpha and PDGF R beta are members of the class III subfamily of RTK which are characterized by the presence of five immunoglobulin-like domains in their extracellular region and a split kinase domain in their intracellular region. PDGF binding induces receptor homo- and heterodimerization leading to signal transduction. The expression of PDGF alpha and beta receptors is independently regulated in various cell types.
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Entrez Gene IDs:
5159 (Human); 18596 (Mouse); 24629 (Rat);
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Long Name:
Platelet-derived Growth Factor Receptor beta
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Aliases:
beta-type platelet-derived growth factor receptor; CD140 antigen-like family member B; CD140b antigen; CD140b; EC 2.7.10; EC 2.7.10.1; JTK12; PDGF R beta; PDGFR; PDGFR1; PDGFRB; PDGF-R-beta; platelet-derived growth factor receptor, beta polypeptide