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.
The BMP-9 BioAssay™ ELISA Kit is a Sandwich High Sensitivity ELISA for quantitative detection of human BMP-9 in bone tissue, cell culture supernates, serum, plasma (heparin, EDTA) and urine.
Detection Range:15.6pg/ml-1000pg/ml
Sensitivity:<10pg/ml
Specificity:Recognizes natural and recombinant Human BMP-9. There is no detectable cross-reactivity with other relevant proteins.
Test Principle:This Human BMP-9 BioAssay™ ELISA Kit is a solid phase immunoassay specially designed to measure human BMP-9 with a 96-well strip plate that is pre-coated with antibody specific for BMP-9. The detection antibody is abiotinylated antibody specific for BMP-9. The capture antibody is a monoclonal antibody from mouse, the detection antibody is a biotinylatedpolyclonal antibody from goat. The kit contains recombinant human BMP-9 with immunogen: Expression system for standard: CHO, Immunogensequence: S320-R429. To measure human BMP-9, add standards and samples to the wells, then add the biotinylated detection antibody. Wash the wells with PBS or TBSbuffer, and add Avidin-Biotin-Peroxidase Complex (ABC-HRP). Wash away the unbounded ABC-HRP with PBS or TBS buffer and add TMB. TMBis substrate to HRP and will be catalyzed to produce a blue color product, which changes into yellow after adding acidic stop solution. The densityof the yellow product is linearly propotional to human BMP-9 in the sample. Read the density of the yellow product in each well using a platereader, and benchmark the sample wells' readings against the standard curve to determine the concentration of human BMP-9 in the sample.
Storage and Stability:Store *396590B powder at 4°C. Once reconstituted store at 4°C for up to 12 hours or at -20°C for up to 48 hours. Store other components at 4°C. Stable for 6 months after receipt. For maximum recovery of product, centrifuge the original vial after thawing and prior to removing the cap.
Assay Summary:1. Add 100ul of samples and standards and incubate the plate at 37°C for 90 minutes or at RT for 2 hours. Do not wash
2. Add 100ul biotinylated antibody and incubate the plate at 37°C for 60 minutes or at RT for 90 minutes. Wash plate 3 times with 0.01M PBS or TBS
3. Add 100ul of ABC working solution and incubate the plate at 37°C for 30 minutes or at RT for 40 minutes. Wash plate 5 times with 0.01M PBS or TBS
4. Add 90ul of TMB color developing agent and incubate the plate at 37°C in dark for 15-25 minutes or at RT for 30 minutes
