S100A8 (also known as MRP8, Calgranulin A, and CP-10) and S100A9 (also known as MRP14 and Calgranulin B) are secreted pro-inflammatory proteins that are upregulated in neutrophils and monocytes at sites of inflammation (e.g. psoriasis, rheumatoid arthritis, cardiac ischemia) and are present at elevated concentrations in rheumatoid arthritis synovial fluid. In the presence of calcium or zinc, S100A8 and S100A9 associate into noncovalent homodimers and heterodimers with each other. The heterodimer additionally binds and sequesters manganese, thereby restricting the growth of Mn-dependent bacteria. The S100A8/A9 heterodimer exhibits functions beyond those performed by the individual proteins. These include binding to fatty acids such as arachidonic acid and promoting astrocyte proliferation. S100A8, S100A9, and the heterodimer each promote neutrophil infiltration into sites of inflammation and inflammatory cytokine production by monocytes.
Mouse S100A8/S100A9 Heterodimer DuoSet ELISA
R&D Systems | Catalog # DY8596-05
Key Product Details
Assay Type
Assay Range
Sample Type
Note: Diluents for complex matrices, such as serum and plasma, should be evaluated prior to use in this DuoSet
Reactivity
Mouse S100A8/S100A9 Heterodimer DuoSet ELISA 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
- Economical alternative to complete kits
Product Summary for Mouse S100A8/S100A9 Heterodimer DuoSet ELISA
This DuoSet ELISA Development kit contains the basic components required for the development of sandwich ELISAs to measure natural and Recombinant mouse S100 Calcium Binding Protein A8 and S100 Calcium Binding Protein A9 Heterodimer (S100A8/S100A9 Heterodimer). The suggested diluent is suitable for the analysis of most cell culture supernate, serum, and plasma samples. Diluents for complex matrices, such as serum and plasma, should be evaluated prior to use in this DuoSet.
Product Specifications
Assay Format
Detection Method
Conjugate
Specificity
Label
Scientific Data Images for Mouse S100A8/S100A9 Heterodimer DuoSet ELISA
Mouse S100A8 / S100A9 Heterodimer ELISA Standard Curve
Kit Contents for Mouse S100A8/S100A9 Heterodimer DuoSet ELISA
- Capture Antibody
- Detection Antibody
- Recombinant Standard
- Streptavidin conjugated to horseradish-peroxidase (Streptavidin-HRP)
Other Reagents Required
PBS: (Catalog # DY006), or 137 mM NaCl, 2.7 mM KCl, 8.1 mM Na2HPO4, 1.5 mM KH2PO4, pH 7.2 - 7.4, 0.2 µm filtered
Wash Buffer: (Catalog # WA126), or equivalent
Reagent Diluent*
Blocking Buffer*
Substrate Solution: ELISA TMB Substrate (Catalog # DY999B or DY999B-250)
Stop Solution: Methanesulfonic acid (Catalog # DY994B or DY994B-250)
Microplates: (Catalog # DY990), or equivalent
Plate Sealers: (Catalog # DY992), or equivalent
*For the recommended Reagent Diluent and Blocking Buffer for a specific DuoSet ELISA Development Kit, refer to the product datasheet.
Preparation and Storage
Shipping
Stability & Storage
Background: S100A8/S100A9 Heterodimer
Long Name
Alternate Names
Entrez Gene IDs
Gene Symbol
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Product Specific Notices for Mouse S100A8/S100A9 Heterodimer DuoSet ELISA
For research use only
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Protocols
View specific protocols for Mouse S100A8/S100A9 Heterodimer DuoSet ELISA (DY8596-05):
1. Dilute the Capture Antibody to the working concentration in PBS without carrier protein. Immediately coat a 96-well microplate with 100 μL per well of the diluted Capture Antibody. Seal the plate and incubate overnight at room temperature.
2. Aspirate each well and wash with Wash Buffer, repeating the process two times for a total of three washes. Wash by filling each well with Wash Buffer (400 μL) using a squirt bottle, manifold dispenser, or autowasher. Complete removal of liquid at each step is essential for good performance. After the last wash, remove any remaining Wash Buffer by aspirating or by inverting the plate and blotting it against clean paper towels.
3. Block plates by adding 300 μL of Block Buffer to each well. Incubate at room temperature for a minimum of 1 hour.
4. Repeat the aspiration/wash as in step 2. The plates are now ready for sample addition.
Assay Procedure
1. Add 100 μL of sample or standards in Reagent Diluent, or an appropriate diluent, per well. Cover with an adhesive strip and incubate 2 hours at room temperature.
2. Repeat the aspiration/wash as in step 2 of Plate Preparation.
3. Add 100 μL of the Detection Antibody, diluted in Reagent Diluent, to each well. Cover with a new adhesive strip and incubate 2 hours at room temperature.
4. Repeat the aspiration/wash as in step 2 of Plate Preparation.
5. Add 100 μL of the working dilution of Streptavidin-HRP to each well. Cover the plate and incubate for 20 minutes at room temperature. Avoid placing the plate in direct light.
6. Repeat the aspiration/wash as in step 2.
7. Add 100 μL of Substrate Solution to each well. Incubate for
20 minutes at room temperature. Avoid placing the plate in direct light.
8. Add 50 μL of Stop Solution to each well. Gently tap the plate to ensure thorough mixing.
9. Determine the optical density of each well immediately, using a microplate reader set to 450 nm. If wavelength correction is available, set to 540 nm or 570 nm. If wavelength correction is not available, subtract readings at 540 nm or 570 nm from the readings at 450 nm. This subtraction will correct for optical imperfections in the plate. Readings made directly at 450 nm without correction may be higher and less accurate.
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