• Assay Type

  Solid Phase Sandwich ELISA

• Format

  96-well strip plate

• Range

  93.80 - 6,000 pg/mL

• Sufficient Materials

  For five 96-well plates*

• 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 ELISA Development kit contains the basic components required for the development of sandwich ELISAs to measure natural and Recombinant human S100 Calcium Binding Protein A8/S100 Calcium Binding Protein A9 (S100A8/S100A9 Heterodimer). The suggested diluent is suitable for the analysis of most cell culture supernate, serum, plasma, saliva, human milk, urine, and fecal samples. Diluents for complex matrices, such as serum, plasma, saliva, human milk, urine, and feces, 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 # ), or 137 mM NaCl, 2.7 mM KCl, 8.1 mM Na  ₂HPO  ₄, 1.5 mM KH  ₂PO  ₄, pH 7.2 - 7.4, 0.2 µm filtered  
 
  Wash Buffer: (Catalog # ), or equivalent  
 
  Reagent 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: R&D Systems (Catalog # ), or equivalent  
 
  Plate Sealers: ELISA Plate Sealers (Catalog # ), or equivalent  
 
  Normal Goat Serum: R&D Systems (Catalog # ), or equivalent

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

Preparation and Storage

• 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 Ca  ²⁺-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 Ca  ²⁺-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.

• Entrez Gene IDs:

  6279 (Human);

• Long Name:

  S100 Calcium Binding Protein A8/S100 Calcium Binding Protein A9

• Aliases:

  Calprotectin; S100A8/S100A9 Heterodimer