Growth/differentiation factor 5 is a protein that in humans is encoded by the GDF5 gene. GDF5 is a protein belonging to the transforming growth factor beta superfamily that is expressed in the developing central nervous system, and has a role in skeletal and joint development. It also increases the survival of neurones that respond to the neurotransmitter dopamine, and is a potential therapeutic molecule associated with Parkinsons disease. 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 Rat GDF5/Bmp 14. Sensitivity: <10pg/ml Range: 31.2pg/ml-2000pg/ml Specificity: Recognizes natural and recombinant Rat GDF5/Bmp 14. There is no detectable cross-reactivity with other relevant proteins. Test Principle: This Rat GDF5/Bmp 14 BioAssay(TM) ELISA Kit was based on standard sandwich enzyme-linked immune- sorbent assay technology. A specific for GDF5/Bmp 14 has been precoated onto 96-well plates. Standards(Expression system for standard: E.coli, Immunogen sequence: A376-R495) and test samples are added to the wells, a biotinylated detection specific for GDF5/Bmp 14 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 GDF5/Bmp 14 amount of sample captured in plate. Kit Components: 96-well plate precoated with anti-Rat Gdf5 antibody, 1x plate Lyophilized recombinant Rat Gdf5 standard, 2x10ng/tube Biotinylated anti-Rat Gdf5 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.
