Sclerostin is a protein that in humans is encoded by the SOST gene. It is a secreted glycoprotein with a C-terminal cysteine knot-like (CTCK) domain and sequence similarity to the DAN (differential screening-selected gene aberrative in neuroblastoma) family of bone morphogenetic protein (BMP) antagonists. Additionally, Sclerostin is produced by the osteocyte and has anti-anabolic effects on bone formation. The SOST gene maps to chromosome 17q12-q21. 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 Human Sclerostin/SOST. Sensitivity: <10pg/ml Range: 31.2pg/ml-2000pg/ml Specificity: Recognizes natural and recombinant Human Sclerostin/SOST. There is no detectable cross-reactivity with other relevant proteins. Test Principle: This Human Sclerostin/SOST BioAssay(TM) ELISA Kit was based on standard sandwich enzyme-linked immune- sorbent assay technology. A monoclonal antibody from mouse specific for Sclerostin/SOST has been precoated onto 96-well plates. Standards(Expression system for standard: NSO, Immunogen sequence: Q24-Y213) and test samples are added to the wells, a biotinylated detection polyclonal antibody from goat specific for Sclerostin/SOST 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 Sclerostin/SOST amount of sample captured in plate. Kit Components: 96-well plate precoated with anti-Human SOST antibody, 1x plate Lyophilized recombinant Human SOST standard, 2x10ng/tube Biotinylated anti-Human SOST 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.
