Growth differentiation factor 2 (GDF2) also known as bone morphogenetic protein (BMP)-9 is a secreted ligand of the TGF-beta (transforming growth factor-beta) superfamily of proteins. Ligands of this family bind various TGF-beta receptors leading to recruitment and activation of SMAD family transcription factors that regulate gene expression. The preproprotein is proteolytically processed to generate each subunit of the disulfide-linked homodimer. This protein regulates cartilage and bone development, angiogenesis and differentiation of cholinergic central nervous system neurons. Homozygous null mice exhibit malformations of the vasculature and skeleton. Sample Type: Cell culture supernates, serum, bone tissue, tissue, plasma(Heparin or EDTA). Intended Use: Sandwich High Sensitivity BioAssay(TM) ELISA kit for Quantitative Detection of Rat BMP-9. Sensitivity: <10pg/ml Range: 15.6pg/ml-1000pg/ml Specificity: Recognizes natural and recombinant Rat BMP-9. There is no detectable cross-reactivity with other relevant proteins. Test Principle: This Rat BMP-9 BioAssay(TM) ELISA Kit was based on standard sandwich enzyme-linked immune-sorbent assay technology. A a monoclonal antibody from mouse specific for BMP-9 has been precoated onto 96-well plates. Standards(Expression system for standard: CHO, Immunogen sequence: S320-R430) and test samples are added to the wells, a biotinylated detection a biotinylated polyclonal antibody from goat specific for BMP-9 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 BMP-9 amount of sample captured in plate. Kit Components: 96-well plate precoated with anti-Rat Gdf2 antibody, 1x plate Lyophilized recombinant Rat Gdf2 standard, 2x10ng/tube Biotinylated anti-Rat Gdf2 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.
