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. Sample Type: Cell lysates, cell culture supernates, serum, plasma (Heparin or EDTA). Intended Use: Sandwich High Sensitivity BioAssay(TM) ELISA kit for Quantitative Detection of Mouse DKK4. Sensitivity: <10pg/ml Range: 78pg/ml-5000pg/ml Specificity: Recognizes natural and recombinant Mouse DKK4. There is no detectable cross-reactivity with other relevant proteins. Test Principle: This Mouse DKK4 BioAssay(TM) ELISA Kit was based on standard sandwich enzyme-linked immune-sorbent assay technology. A 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 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. Kit Components: 96-well plate precoated with anti-Mouse Dkk4 antibody, 1x plate Lyophilized recombinant Mouse Dkk4 standard, 2x10ng/tube Biotinylated anti-Mouse Dkk4 antibody, 1x130ul (dilution 1:100) Avidin-Biotin-Peroxidase Complex (ABC), 1x130ul (dilution 1:100) Sample diluent buffer, 1x30ml Antibody diluent buffer, 1x12ml ABC diluent buffer, 1x12ml TMB color developing agent, 1x10ml TMB stop solution, 1x10ml PBS washing buffer Powder for 1000ml, 1x buffer Storage and Stability: Store powder at 4C liquid at -20C. Store other components at 4C. Stable for at least 6 months For maximum recovery of product, centrifuge the original vial after thawing and prior to removing the cap.
