Dickkopf-related protein 3 is a protein that in humans is encoded by the DKK3 gene. This gene encodes a protein that is a member of the dickkopf family. It is mapped to 11p15.3. The secreted protein contains two cysteine rich regions and is involved in embryonic development through its interactions with the Wnt signaling pathway. The expression of this gene is decreased in a variety of cancer cell lines and it may function as a tumor suppressor gene. Members of the Dkk-related family display unique patterns of mRNA expression in human and mouse tissues, and are secreted when expressed in 293T cells. DKKs play an important role in vertebrate development, where they locally inhibit Wnt regulated processes such as antero-posterior axial patterning, limb development, somitogenesis and eye formation. In the adult, Dkks are implicated in bone formation and bone disease, cancer and Alzheimer disease.
Sample Type:Cell culture supernates, serum and plasma (heparin, EDTA)
Intended Use:For quantitative detection of human DKK-3 in cell culture supernates, serum and plasma (heparin, EDTA).
Sensitivity:<10pg/ml
Range:156-10,000pg/ml
Specificity:Natural and recombinant human DKK-3
Test Principle:This human DKK-3 BioAssay™ ELISA Kit was based on standard sandwich enzyme-linked immune-sorbent assay technology. A monoclonal antibody from mouse specific for DKK-3 has been precoated onto 96-well plates. Standards(NSO, A22-I350) and test samples are added to the wells, a biotinylated detection polyclonal antibody from goat specific for DKK-3 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 DKK-3 amount of sample captured in plate.
Kit Components:96-well plate precoated with human DKK-3 antibody, 1Lyophilized recombinant human DKK-3 standard, 10ng/tube×2Biotinylated human DKK-3 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.
