S100 calcium-binding protein A8 (S100A8), also known as calgranulin A, is a protein that in humans is encoded by the S100A8 gene. It is mapped to chromosome 1q21.3 based on an alignment of the S100A8 sequence with the genomic sequence. The proteins S100A8 and S100A9 form a heterodimer called calprotectin which is a major calcium- and zinc-binding protein in the cytosol of neutrophils, monocytes, and keratinocytes. This gene is a member of the S100 family of proteins containing 2 EF- hand calcium-binding motifs. S100 proteins are localized in the cytoplasm and/or nucleus of a wide range of cells, and involved in the regulation of a number of cellular processes such as cell cycle progression and differentiation. And S100 genes include at least 13 members which are located as a cluster on chromosome 1q21. This protein may function in the inhibition of casein kinase and as a cytokine. Altered expression of this protein is associated with the disease cystic fibrosis. Sample Type: Cell lysates, cell culture supernates, serum, plasma (Heparin or EDTA). Intended Use: Sandwich High Sensitivity BioAssay(TM) ELISA kit for Quantitative Detection of Human S100A8/Calgranulin A. Sensitivity: <10pg/ml Range: 31.2pg/ml-2000pg/ml Specificity: Recognizes natural and recombinant Human S100A8/Calgranulin A. There is no detectable cross-reactivity with other relevant proteins. Test Principle: This Human S100A8/Calgranulin A BioAssay(TM) ELISA Kit was based on standard sandwich enzyme-linked immune-sorbent assay technology. A specific for S100A8/Calgranulin A has been precoated onto 96-well plates. Standards(Expression system for standard: E.coli, Immunogen sequence: L2-E93) and test samples are added to the wells, a biotinylated detection specific for S100A8/Calgranulin A is added subsequently and then followed by washing with PBS or TBS buffer. Avidin-Biotin-Peroxidase Complex was added and unbound conjugates were washed away with PBS or TBS buffer. HRP substrate TMB was used to visualize HRP enzymatic reaction. TMB was catalyzed by HRP to produce a blue color product that changed into yellow after adding acidic stop solution. The density of yellow is proportional to the Human S100A8/Calgranulin A amount of sample captured in plate. Kit Components: 96-well plate precoated with anti-Human S100A8 antibody, 1x plate Lyophilized recombinant Human S100A8 standard, 2x2ng/tube Biotinylated anti-Human S100A8 antibody, 1x130ul (dilution 1:100) Avidin-Biotin-Peroxidase Complex (ABC), 1x130ul (dilution 1:100) Sample diluent buffer, 1x30ml Antibody diluent buffer, 1x12ml ABC diluent buffer, 1x12ml TMB color developing agent, 1x10ml TMB stop solution, 1x10ml PBS washing buffer Powder for 1000ml, 1x buffer Storage and Stability: Store powder at 4C liquid at -20C. Store other components at 4C. Stable for at least 6 months For maximum recovery of product, centrifuge the original vial after thawing and prior to removing the cap.
