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 ELISA kit for Quantitative Detection of Rat Sclerostin/SOST.Sensitivity:<10pg/mlRange:15.6pg/ml-1000pg/mlSpecificity:Recognizes natural and recombinant Rat Sclerostin/SOST. There is no detectable cross-reactivity with other relevant proteins.Test Principle:This Rat Sclerostin/SOST BioAssay ELISA Kit was based on standard sandwich enzyme-linked immune- sorbent assay technology. A specific for Sclerostin/SOST has been precoated onto 96-well plates. Standards(Expression system for standard: NSO; Immunogen sequence: F29-Y213) and test samples are added to the wells, a biotinylated detection 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 Rat Sclerostin/SOST amount of sample captured in plate.Kit Components:96-well plate precoated with anti-Rat Sost antibody, 1x plateLyophilized recombinant Rat Sost standard, 2x10ng/tubeBiotinylated anti-Rat Sost antibody, 1x130ul (dilution 1:100)Avidin-Biotin-Peroxidase Complex (ABC), 1x130ul (dilution 1:100)Sample diluent buffer, 1x30mlAntibody diluent buffer, 1x12mlABC diluent buffer, 1x12mlTMB color developing agent, 1x10mlTMB stop solution, 1x10mlPBS washing buffer Powder for 1000ml, 1x bufferStorage 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.