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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: Insulin R/CD220
Insulin Receptor (INSR, CD220) is a type I transmembrane glycoprotein in the Insulin/IGF Receptor family of receptor tyrosine kinases. The mature receptor contains an extracellular alpha subunit and an extracellular/transmembrane/cytoplasmic beta subunit. Alternative splicing generates A and B isoforms which may homodimerize or heterodimerize with the IGF-I receptor. All receptor combinations bind Insulin, IGF-I, or IGF-II, but with differing affinities. Insulin R signaling regulates glucose uptake and metabolism but also contributes to cell growth, differentiation and apoptosis. Mutations in INSR have been linked to severe insulin resistance (type A and Rabson-Mendenhall syndrome), type II diabetes mellitus, and leprechaunism (Donohue syndrome).