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Assay Type
Solid Phase Sandwich ELISA
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Format
96-well strip plate
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Assay Length
4 hours 40 minutes (after plate preparation)
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Sample Type & Volume Required
Cell lysates (100 µL)
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Range
312.50 - 20,000 pg/mL
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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 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 with 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
- Streptavidin-HRP
Other Reagents Required
PBS: (Catalog # ), or 137 mM NaCl, 2.7 mM KCl, 8.1 mM Na
2HPO
4, 1.5 mM KH
2O
4, 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 (H
2O
2) and Color Reagent B (Tetramethylbenzidine) (Catalog # )
Stop Solution: 2 N H
2SO
4 (Catalog # )
Microplates: From Costar EIA Plate (Costar Catalog # 2592) or 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: Ribosomal Protein S6/RPS6
40S Ribosomal Protein S6 (RPS6) is the major substrate of protein kinases, particularly p70 S6 kinase, in eukaryotic ribosomes. RPS6 phosphorylation at S235, S236, S240, and S244 upregulates the translation of mRNAs containing an oligopyrimidine tract at their transcriptional start sites. This phosphorylation of RPS6 is stimulated by growth factors, tumor promoting agents, and other mitogens. RPS6 is dephosphorylated during growth arrest.
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Entrez Gene IDs:
6194 (Human); 20104 (Mouse); 29304 (Rat);
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Aliases:
Phosphoprotein NP33, 40S ribosomal protein S6; pS6; Ribosomal Protein S6; RPS6; S6
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