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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 mins (after plate preparation)
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Sample Type & Volume Required
Cell lysates (100 µL)
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Range
156.00 - 10,000 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 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 both phosphorylated and unphosphorylated protein, utilizing a standard Streptavidin-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: HO-1/HMOX1/HSP32
Heme Oxygenase (HO), also known as HMOX, is the rate limiting enzyme in the breakdown of heme into Biliverdin, CO, and iron. There are two major isoforms of this enzyme, HO-1 and HO-2. Both have similar enzymatic activity; however, HO-1 is rapidly inducible, while HO-2 is constitutively expressed. Inducers of HO-1 include cell stressors such as oxidative stress, infection, hypoxia, and cytotoxic agents. The reaction products generated by HO-1 activity are biologically active, and the enzyme has a broad range of putative roles. For instance, Biliverdin is an antioxidant with cytoprotective and anti-inflammatory properties, and CO has the potential to suppress inflammation and apoptosis. Because of these activities, HO-1 has received much attention as a therapeutic target for a range of pathological processes.
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Entrez Gene IDs:
3162 (Human); 24451 (Mouse); 15368 (Rat);
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Long Name:
Heme Oxygenase 1
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Aliases:
bK286B10; EC 1.14.99.3; heat shock protein, 32-kD; heme oxygenase (decycling) 1; HMOX1; HO; HO1; HO-1; HO-1heme oxygenase 1; HSP32