Mouse S100A8/S100A9 Heterodimer DuoSet ELISA
Mouse S100A8/S100A9 Heterodimer DuoSet ELISA Summary
* 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
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
Product Datasheets
Preparation and Storage
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.
Assay Procedure
Plate Preparation1. 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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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
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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
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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
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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
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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
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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
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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
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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
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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
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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
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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
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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
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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
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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
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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
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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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