Nidogen-1(NID-1), also known as Entactin, is a protein that in humans is encoded by the NID1 gene. It is a member of the nidogen family of basement membrane glycoproteins. This gene is mapped to 1q42.3. Nidogen-1 is a component of the basement membrane alongside other components such as collagen type IV, proteoglycans(heparan sulfate and glycosaminoglycans), laminin and fibronectin. The protein interacts with several other components of basement membranes. Structurally it(along with perlecan) connects the networks formed by collagens and laminins to each other. It may also play a role in cell interactions with the extracellular matrix. Nidogen-1 also can serve as a bridge between the 2 most abundant molecules in the basement membrane: type IV collagen and laminin.
Sample Type:Cell culture supernates and serum.
Intended Use:For quantitative detection of human Nidogen-1 in cell culture supernates and serum.
Sensitivity:<10pg/ml
Range:62.5-4000pg/ml
Specificity:Natural and recombinant human Nidogen-1
Test Principle:This human Nidogen-1 BioAssay™ ELISA Kit was based on standard sandwich enzyme-linked immune-sorbent assay technology. A monoclonal antibody from mouse specific for Nidogen-1 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 Nidogen-1 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 Nidogen-1 amount of sample captured in plate.
Kit Components:96-well plate precoated with human Nidogen-1 antibody, 1Lyophilized recombinant human Nidogen-1 standard, 10ng/tube×2Biotinylated human Nidogen-1 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.
