Description
For quantitative detection of mouse DKK4 in cell culture supernates, cell lysates, tissue homogenates, serum and plasma (heparin, EDTA).
Background: DKK4 is a membersof the Dickkopf family which are negative regulators of the growth-promoting Wnt / beta-catenin signaling pathway. By FISH, the DKK4 gene was mapped to chromosome 8p11.2-p11.1. The secreted protein contains two cysteine rich regions and is involved in embryonic development through its interactions with the Wnt signaling pathway. Activity of this protein is modulated by binding to the Wnt co-receptor and the co-factor kremen 2. The expression of DKK4 was significantly reduced in hepatocellular carcinomas (HCCs), particularly in HCCs that showed cytoplasmic beta- catenin accumulation. DKK4 expression was also reduced in HCC cell lines. Ectopic expression of DKK4 in 2 HCC cell lines reduced beta-catenin and cyclin D1 protein levels and attenuated beta-catenin activity in reporter gene assays.
Principle: Mouse DKK4 ELISA Kit was based on standard sandwich enzyme-linked immune-sorbent assay technology. A monoclonal antibody from Rat specific for DKK4 has been precoated onto 96-well plates. Standards (Expression system for standard: NSO, Immunogen sequence: L19-I221) and test samples are added to the wells, a biotinylated detection polyclonal antibody from goat specific for DKK4 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 Mouse DKK4 amount of sample captured in plate