Nectin-4, also known as PVRL4, is an adhesion receptor of the immunoglobulin superfamily and functions in cell-cell adhesion. This gene is mapped to chromosome 1q23.3. The encoded protein contains two immunoglobulin-like (Ig-like) C2-type domains and one Ig-like V-type domain. It is involved in cell adhesion through trans-homophilic and -heterophilic interactions. It is a single-pass type I membrane protein. The soluble form is produced by proteolytic cleavage at the cell surface by the metalloproteinase ADAM17/TACE. The secreted form is found in both breast tumor cell lines and breast tumor patients. Mutations in this gene are the cause of ectodermal dysplasia-syndactyly syndrome type 1, an autosomal recessive disorder. Alternatively spliced transcript variants have been found but the full-length nature of the variant has not been determined. Sample Type: Cell lysate, cell culture supernates, serum, urine, plasma (Heparin or EDTA). Intended Use: Sandwich High Sensitivity BioAssay(TM) ELISA kit for Quantitative Detection of Human Nectin-4/PVRL4. Sensitivity: <10pg/ml Range: 31.2pg/ml-2000pg/ml Specificity: Recognizes natural and recombinant Human Nectin-4/PVRL4. There is no detectable cross-reactivity with other relevant proteins. Test Principle: This Human Nectin-4/PVRL4 BioAssay(TM) ELISA Kit was based on standard sandwich enzyme-linked immune- sorbent assay technology. A specific for Nectin-4/PVRL4 has been precoated onto 96-well plates. Standards(Expression system for standard: NSO, Immunogen sequence: G32-S349) and test samples are added to the wells, a biotinylated detection specific for Nectin-4/PVRL4 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 Nectin-4/PVRL4 amount of sample captured in plate. Kit Components: 96-well plate precoated with anti-Human NECTIN4 antibody, 1x plate Lyophilized recombinant Human NECTIN4 standard, 2x10ng/tube Biotinylated anti-Human NECTIN4 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.
