CD47 (Cluster of Differentiation 47), also known as integrin associated protein (IAP), is a transmembrane protein that in humans is encoded by the CD47 gene. It belongs to the immunoglobulin superfamily and partners with membrane integrins and also binds the ligands thrombospondin-1 (TSP-1) and signal-regulatory protein alpha (SIRPalpha). CD47 is involved in a range of cellular processes, including apoptosis, proliferation, adhesion, and migration. Furthermore, it plays a key role in immune and angiogenic responses. Also CD47 is ubiquitously expressed in human cells and has found to be overexpressed in many different tumor cells.
Sample Type:Cell culture supernates and serum.
Intended Use:For quantitative detection of human CD47 in cell culture supernates and serum.
Sensitivity:<10pg/ml
Range:125-8000pg/ml
Specificity:Natural and recombinant human CD47
Test Principle:This human CD47 BioAssay™ ELISA Kit was based on standard sandwich enzyme-linked immune-sorbent assay technology. A monoclonal antibody from mouse specific for CD47 has been precoated onto 96-well plates. Standards (NSO, Q19 - P139) and test samples are added to the wells, a biotinylated detection polyclonal antibody from goat specific for CD47 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 CD47 amount of sample captured in plate.
Kit Components:96-well plate precoated with human CD47 antibody, 1Lyophilized recombinant human CD47 standard, 10ng/tube×2Biotinylated human CD47 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.
