Sialic acid-binding Ig-like lectin 5 is a protein that in humans is encoded by the SIGLEC5 gene. This gene encodes a member of the sialic acid-binding immunoglobulin-like lectin (Siglec) family. These cell surface lectins are characterized by structural motifs in the immunoglobulin (Ig)-like domains and sialic acid recognition sites in the first Ig V set domain. The encoded protein is a member of the CD33-related subset of Siglecs and inhibits the activation of several cell types including monocytes, macrophages and neutrophils. Binding of group B Streptococcus (GBS) to the encoded protein plays a role in GBS immune evasion. 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 Siglec5/Cd170. Sensitivity: <15pg/ml Range: 46.9pg/ml-3000pg/ml Specificity: Recognizes natural and recombinant Human Siglec5/Cd170. There is no detectable cross-reactivity with other relevant proteins. Test Principle: This Human Siglec5/Cd170 BioAssay(TM) ELISA Kit was based on standard sandwich enzyme-linked immune- sorbent assay technology. A monoclonal antibody from mouse specific for Siglec5/Cd170 has been precoated onto 96-well plates. Standards(Expression system for standard: NSO, Immunogen sequence: E17-L441) and test samples are added to the wells, a biotinylated detection polyclonal antibody from goat specific for Siglec5/Cd170 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 Siglec5/Cd170 amount of sample captured in plate. Kit Components: 96-well plate precoated with anti-Human SIGLEC5 antibody, 1x plate Lyophilized recombinant Human SIGLEC5 standard, 2x10ng/tube Biotinylated anti-Human SIGLEC5 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.
