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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, an HRP-conjugated 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 alpha
PDGF R alpha (Platelet-Derived Growth Factor Receptor alpha) is a transmembrane receptor tyrosine kinase that can form homodimers or heterodimers with PDGF R beta when engaged by dimers of the PDGF ligands. PDGF R alpha binds to PDGF-AA, PDGF-AB, and PDGF-CC. It is strongly expressed in oligodendrocyte, lung, skin, and intestinal progenitor cells, at low levels in most mesenchymal cells, and can be induced by inflammation. PDGF R alpha function is important during formation of the cranial and cardiac neural crest, retina, gonads, lung alveoli, intestinal villi, skin, hair follicles, skeleton, teeth, palate, and interstitial kidney mesenchyme. Dysregulated PDGF R alpha signaling can result in spina bifida or cleft palate-type malformations. Postnatally, PDGF R alpha is implicated in gliomas and fibrotic disorders of lung, heart, and skin (scleroderma).
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Entrez Gene IDs:
5156 (Human); 18595 (Mouse);
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Long Name:
Platelet-derived Growth Factor Receptor alpha
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Aliases:
alpha-type platelet-derived growth factor receptor; CD140 antigen-like family member A; CD140a antigen; CD140a; EC 2.7.10; EC 2.7.10.1; MGC74795; PDGF R alpha; PDGFR2; PDGFRA; PDGFRA/BCR fusion; PDGF-R-alpha; platelet-derived growth factor receptor, alpha polypeptide; rearranged-in-hypereosinophilia-platelet derived growth factor receptor alphafusion protein; RHEPDGFRA