The protein CD80 (Cluster of Differentiation 80) is a molecule found on activated B cells and monocytes which provides a costimulatory signal necessary for T cell activation and survival. It is also known as B7-1. The cDNA for B7-1 predicts a type I membrane protein, i.e., one synthesized with a signal peptide that is cleaved upon translocation across the endoplasmic membrane. The protein is predicted to contain 2 extracellular domains structurally similar to those of Ig, a hydrophobic transmembrane region, and a short cytoplasmic domain.The CD80 and CD86 genes encode B7-1 and B7-2, respectively, which are structurally similar members of the immunoglobulin superfamily expressed on a variety of hematopoietic cell types. B7-1 and B7-2 provide a costimulatory signal to T cells by interacting with CD28 and CTLA4.
Sample Type:Cell culture supernates and serum.
Intended Use:For quantitative detection of rat CD80 in cell culture supernates and serum.
Sensitivity:<2pg/ml
Range:62.5-4000pg/ml
Specificity:Natural and recombinant rat CD80
Test Principle:This rat CD80 BioAssay™ ELISA Kit was based on standard sandwich enzyme-linked immune-sorbent assay technology. A monoclonal antibody from mouse specific for CD80 has been precoated onto 96-well plates. Standards(NSO) and test samples are added to the wells, a biotinylated detection polyclonal antibody from goat specific for CD80 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 rat CD80 amount of sample captured in plate.
Kit Components:96-well plate precoated with rat CD80 antibody, 1Lyophilized recombinant rat CD80 standard, 10ng/tube×2Biotinylated rat CD80 antibody, 130ul (dilution 1:100)Avidin-Biotin-Peroxidase Complex (ABC), 130ul (dilution 1:100)Sample diluent buffer, 30mlAntibody diluent buffer, 12mlABC diluent buffer, 12mlTMB color developing agent, 10mlTMB stop solution, 10ml
Storage and Stability:Store powder at 4°C liquid at -20°C. Store other components at 4°C. Stable for at least 6 months For maximum recovery of product, centrifuge the original vial after thawing and prior to removing the cap.
