Description
Key facts
Detection method:Colorimetric,
Sample types:Plasma, Cell culture supernatant, Serum,
Reacts with:Human,
Assay type:Sandwich (quantitative),
Sensitivity:= 35 pg/mL,
Range:32.77 - 8000 pg/mL,
Assay Platform:Pre-coated microplate (12 x 8 well strips)
Product details:
Human Calprotectin ELISA kit (S100A8/S100A9) ab267628 is a sandwich ELISA to measure Human Calprotectin (S100A8/S100A9) in serum, plasma, cell culture supernatant with a sensitivity of 35 pg/ml.
How the assay works
This assay employs an antibody specific for human Calprotectin coated on a 96-well plate. Standards and samples are pipetted into the wells and Calprotectin present in a sample is bound to the wells by the immobilized antibody. The wells are washed and biotinylated anti-human Calprotectin antibody is added. After washing away unbound biotinylated antibody, HRP-conjugated streptavidin is pipetted to the wells. The wells are again washed, a TMB substrate solution is added to the wells and color develops in proportion to the amount of Calprotectin bound. The Stop Solution changes the color from blue to yellow, and the intensity of the color is measured at 450 nm.
Assay Specificity
Our ELISA kits are rigorously validated to ensure the highest level of consistency and reproducibility. Please check the protocol booklet for more details
Human Calprotectin ELISA kit (S100A8/S100A9) ab267628 protocol summary
1. Add 100 μL standard or sample to each well. Incubate 2.5 hours at room temperature.
Add 100 μL prepared biotin antibody to each well. Incubate 1 hour at room temperature.
Add 100 μL prepared Streptavidin solution. Incubate 45 minutes at room temperature.
2. Add 100 μL TMB One-Step Substrate Reagent to each well. Incubate 30 minutes at room temperature.
3. Add 50 μL Stop Solution to each well. Read at 450 nm immediately.
Properties and Storage Information:
Shipped at conditions-Blue Ice, Appropriate short-term storage conditions--20°C, Appropriate long-term storage conditions--20°C, Storage information--20°C
Supplementary Information:
This supplementary information is collated from multiple sources and compiled automatically.
S100A8 and S100A9 often known together as calprotectin are proteins that form a heterodimer and function as a complex. Each protein has a mass of approximately 10.8 kDa for S100A8 and 13.2 kDa for S100A9. These proteins express mainly in neutrophils and monocytes acting as calcium-binding proteins. Their expression can modulate various cellular functions like migration and inflammation responses.
Biological function summary
Calprotectin serves as an important player in the immune response. This complex possesses antimicrobial properties and contributes to the body's defense by modulating inflammation. It binds to zinc and manganese to deprive pathogens of essential nutrients an action called nutrient bioavailability restriction. In immune cells calprotectin's presence signals the activation and recruitment of further immune components.
Pathways
S100A8 and S100A9 influence the NF-kB signaling pathway which governs immune responses and inflammation. Their function affects other pathways such as MAPK by mediating pro-inflammatory cytokines' production and release. These proteins often engage with Toll-like receptors impacting the innate immune system and contributing to the body's defensive mechanisms against pathogens.
High levels of calprotectin are associated with inflammatory diseases like rheumatoid arthritis and inflammatory bowel disease. Its involvement in these conditions makes it a valuable biomarker. Calprotectin often interacts with other inflammatory molecules like TNF-alpha an important factor in propagating inflammation. The calprotectin ELISA test kit plasma calprotectin immunoassay and similar tools can aid in diagnosing and monitoring these conditions providing insight into disease progression and response to treatment.
