Mouse S100A8/S100A9 Heterodimer DuoSet ELISA

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Mouse S100A8 / S100A9 Heterodimer ELISA Standard Curve
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Product Details
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Mouse S100A8/S100A9 Heterodimer DuoSet ELISA Summary

Assay Type
Solid Phase Sandwich ELISA
96-well strip plate
Assay Range
125.0 - 8,000 pg/mL
Sufficient Materials
For five 96-well plates*
Please see the product datasheet

* 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 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 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

Kit Content

  • 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: 1:1 mixture of Color Reagent A (H2O2) and Color Reagent B (Tetramethylbenzidine) (Catalog # DY999)

Stop Solution: 2 N H2SO4 (Catalog # DY994)

Microplates: R&D Systems (Catalog # DY990), or equivalent

Plate Sealers: ELISA Plate Sealers (Catalog # DY992), or equivalent

*For the Reagent Diluent and Blocking Buffer recommended for a specific DuoSet ELISA Development Kit, please see the product.

Scientific Data

Mouse S100A8 / S100A9 Heterodimer ELISA Standard Curve

Product Datasheets

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Preparation and Storage

The product is shipped at ambient temperature. Upon receipt, store it immediately at the temperature recommended below.
Stability & Storage
Store the unopened product at 2 - 8 °C. Do not use past expiration date.

Background: S100A8/S100A9 Heterodimer

S100A8 (also MRP8 and calgranulin A) is a 10 kDa member of the S100 family, EF-hand superfamily of Ca-binding proteins. It is produced by neutrophils and monocytes, and forms Ca2+-dependent heterodimer/heterotetramer complexes (termed calprotectin) with S100A9. It functions both intracellularly and extracellularly, where it binds to RAGE and CD36. Human S100A8 is 93 amino acids (aa) in length. It contains two EF-hand motifs (aa 12 - 47 and 46 - 81) and one high-affinity Ca2+-binding site (aa 59 - 70). There may be one splice form that shows a 15 aa substitution for the C-terminal 14 amino acids. Although mouse S100A8 is cleaved by MMP-2 after Asn21, it is unclear if human S100A8 is susceptible. Full-length human S100A8 is 57% and 74% aa identical to mouse and canine S100A8, respectively.

Long Name:
S100 Calcium Binding Protein A8/S100 Calcium Binding Protein A9
Entrez Gene IDs:
6279 (Human)
Alternate Names:
Calprotectin; S100A8/S100A9 Heterodimer

Assay Procedure

Plate Preparation
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.

Citations for Mouse S100A8/S100A9 Heterodimer DuoSet ELISA

R&D Systems personnel manually curate a database that contains references using R&D Systems products. The data collected includes not only links to publications in PubMed, but also provides information about sample types, species, and experimental conditions.

16 Citations: Showing 1 - 10
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  1. Neutrophil metalloproteinase driven spleen damage hampers infection control of trypanosomiasis
    Authors: Pham, HTT;Magez, S;Choi, B;Baatar, B;Jung, J;Radwanska, M;
    Nature communications
    Species: Mouse
    Sample Types: Tissue Homogenates
  2. Platelets exacerbate cardiovascular inflammation in a murine model of Kawasaki disease vasculitis
    Authors: Kocatürk, B;Lee, Y;Nosaka, N;Abe, M;Martinon, D;Lane, ME;Moreira, D;Chen, S;Fishbein, MC;Porritt, RA;Franklin, BS;Noval Rivas, M;Arditi, M;
    JCI insight
    Species: Mouse
    Sample Types: Serum
  3. Thrombocytopenia Independently Leads to Monocyte Immune Dysfunction
    Authors: Li, C;Ture, SK;Nieves-Lopez, B;Blick-Nitko, SK;Maurya, P;Livada, AC;Stahl, TJ;Kim, M;Pietropaoli, AP;Morrell, CN;
    bioRxiv : the preprint server for biology
    Species: Mouse
    Sample Types: Plasma
  4. Targeting activin receptor-like kinase 7 ameliorates adiposity and associated metabolic disorders
    Authors: M Zhao, K Okunishi, Y Bu, O Kikuchi, H Wang, T Kitamura, T Izumi
    JCI Insight, 2023-02-22;0(0):.
    Species: Mouse
    Sample Types: Tissue Homogenates
  5. SCCA1/SERPINB3 promotes suppressive immune environment via STAT-dependent chemokine production, blunting the therapy-induced T cell responses
    Authors: L Chen, V Shi, S Wang, R Freeman, F Ruiz, K Jayachandr, J Zhang, P Cosper, L Sun, CJ Luke, C Spina, PW Grigsby, JK Schwarz, S Markovina
    bioRxiv : the preprint server for biology, 2023-02-03;0(0):.
    Species: Mouse
    Sample Types: Tissue Homogenates
  6. Biotin Deficiency Induces Intestinal Dysbiosis Associated with an Inflammatory Bowel Disease-like Phenotype
    Authors: JC Yang, JP Jacobs, M Hwang, S Sabui, F Liang, HM Said, J Skupsky
    Nutrients, 2023-01-04;15(2):.
    Species: Mouse
    Sample Types: Feces
  7. Group B Streptococcus adaptation promotes survival in a hyperinflammatory diabetic wound environment
    Authors: RA Keogh, AL Haeberle, CJ Langouët-A, JS Kavanaugh, EP Schmidt, GD Moore, AR Horswill, KS Doran
    Science Advances, 2022-11-11;8(45):eadd3221.
    Species: Mouse, Transgenic Mouse
    Sample Types: Tissue Homogenates
  8. Keratinocyte-derived cytokine TSLP promotes growth and metastasis of melanoma by regulating the tumor-associated immune microenvironment
    Authors: W Yao, B German, D Chraa, A Braud, C Hugel, P Meyer, G Davidson, P Laurette, G Mengus, E Flatter, P Marschall, J Segaud, M Guivarch, P Hener, MC Birling, D Lipsker, I Davidson, M Li
    JCI Insight, 2022-11-08;0(0):.
    Species: Mouse
    Sample Types: Serum
  9. Pro-inflammatory megakaryocyte gene expression in murine models of breast cancer
    Authors: HG Roweth, MW Malloy, GJ Goreczny, IC Becker, Q Guo, EA Mittendorf, JE Italiano, SS McAllister, EM Battinelli
    Science Advances, 2022-10-12;8(41):eabo5224.
    Species: Mouse
    Sample Types: Cell Culture Supernates
  10. DSS-induced inflammation in the colon drives a proinflammatory signature in the brain that is ameliorated by prophylactic treatment with the S100A9 inhibitor paquinimod
    Authors: S Talley, R Valiauga, L Anderson, AR Cannon, MA Choudhry, EM Campbell
    Journal of Neuroinflammation, 2021-11-10;18(1):263.
    Species: Mouse
    Sample Types: Tissue Homogenates
  11. Geographic differences in gut microbiota composition impact susceptibility to enteric infection
    Authors: AM Porras, Q Shi, H Zhou, R Callahan, G Montenegro, N Solomons, IL Brito
    Cell Reports, 2021-07-27;36(4):109457.
    Species: Mouse
    Sample Types: Stool Supernates
  12. Modeling undernutrition with enteropathy in mice
    Authors: E Salameh, M Jarbeau, FB Morel, M Zeilani, M Aziz, P Déchelotte, R Marion-Let
    Sci Rep, 2020-09-24;10(1):15581.
    Species: Mouse
    Sample Types: Cell Culture Supernates
  13. Transgenic interleukin 11 expression causes cross-tissue fibro-inflammation and an inflammatory bowel phenotype in mice
    Authors: WW Lim, B Ng, A Widjaja, C Xie, L Su, N Ko, SY Lim, XY Kwek, S Lim, SA Cook, S Schafer
    PLoS ONE, 2020-01-09;15(1):e0227505.
    Species: Mouse
    Sample Types: Fecal Extracts
  14. Opposing Effects of IL-1Ra and IL-36Ra on Innate Immune Response to Pseudomonas aeruginosa Infection in C57BL/6 Mouse Corneas
    Authors: N Gao, R Me, C Dai, B Seyoum, FX Yu
    J. Immunol., 2018-06-11;0(0):.
    Species: Mouse
    Sample Types: Tissue Homogenates
  15. Suppression Colitis and Colitis-Associated Colon Cancer by Anti-S100a9 Antibody in Mice
    Authors: X Zhang, L Wei, J Wang, Z Qin, J Wang, Y Lu, X Zheng, Q Peng, Q Ye, F Ai, P Liu, S Wang, G Li, S Shen, J Ma
    Front Immunol, 2017-12-13;8(0):1774.
    Species: Mouse
    Sample Types: Serum
  16. IL-24 Promotes Pseudomonas aeruginosa Keratitis in C57BL/6 Mouse Corneas
    Authors: BX Ross, N Gao, X Cui, TJ Standiford, J Xu, FX Yu
    J. Immunol, 2017-03-22;0(0):.
    Species: Mouse
    Sample Types: Tissue Homogenates


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Mouse S100A8/S100A9 Heterodimer DuoSet ELISA
By Anonymous on 01/27/2024
Sample Tested: n/a,Serum

Mouse S100A8/S100A9 Heterodimer DuoSet ELISA
By Anonymous on 09/13/2021
Sample Tested: Serum