Differential screening-selected gene aberrative in neuroblastoma (DAN) is a member of the DAN family of secreted glycoproteins that are putative BMP antagonists. The NBL1 gene, also known as DAN, is originally cloned from a normal rat fibroblast cDNA library by a differential screening method. The human DAN gene is mapped to chromosome 1p36.13-p36. It is found that the DAN gene possesses a tumor suppressive activity when overexpressed in v-src transformed cells.
Sample Type:Cell culture supernates, serum and plasma (heparin, EDTA)
Intended Use:For quantitative detection of human DAN in cell culture supernates, serum and plasma (heparin, EDTA).
Sensitivity:<10pg/ml
Range:125-8000pg/ml
Specificity:Natural and recombinant human DAN
Test Principle:This human DAN BioAssay™ ELISA Kit was based on standard sandwich enzyme-linked immune-sorbent assay technology. A monoclonal antibody from mouse specific for DAN 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 DAN 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 DAN amount of sample captured in plate.
Kit Components:96-well plate precoated with human DAN antibody, 1Lyophilized recombinant human DAN standard, 10ng/tube×2Biotinylated human DAN 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.
