• Assay Type

  Solid Phase Sandwich ELISA

• Format

  96-well strip plate

• Sample Type & Volume Required

  100 μL

• Range

  31.20 - 2,000 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.

Ancillary Reagent Kit Available  

DY008, DuoSet ELISA Ancillary Reagent Kit 2 -

This DuoSet ELISA Development kit contains the basic components required for the development of sandwich ELISAs to measure natural and Recombinant

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  
 

*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

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); 20201 (Mouse);

• Long Name:

  S100 Calcium Binding Protein A8

• Aliases:

  60B8AG; CAGACP-10; Calgranulin A; calgranulin-A; Calprotectin L1L subunit; CFAG; CFAGL1Ag; CGLA; Cystic fibrosis antigen; Leukocyte L1 complex light chain; MA387; MIF; Migration inhibitory factor-related protein 8; MRP8; MRP-8; MRP8S100 calcium binding protein A8 (calgranulin A); NIF; P8; protein S100-A8; S100 calcium binding protein A8; S100 calcium-binding protein A8 (calgranulin A); S100 calcium-binding protein A8; S100A8; Urinary stone protein band A