Overview

Introduction
Bone morphogenetic protein 4 (BMP-4) is a member of the transforming growth factor-beta superfamily of regulatory molecules. Bone morphogenetic proteins were originally identified by the ability of demineralized bone extract to induce endochondral osteogenesis in vivo in an extraskeletal site, and known for their ability to induce bone and cartilage development. In human embryonic development, BMP4 is a critical signaling molecule required for the early differentiation of the embryo and establishing of a dorsal-ventral axis. BMP4 is secreted from the dorsal portion of the notochord, and it acts in concert with sonic hedgehog (released from the ventral portion of the notochord) to establish a dorsal-ventral axis for the differentiation of later structures. Increase in expression of BMP4 has been associated with a variety of bone diseases, including the heritable disorder Fibrodysplasia Ossificans Progressiva.

Principle of the Assay  
This kit was based on sandwich enzyme-linked immune-sorbent assay technology. Anti- BMP-4 polyclonal antibody was pre-coated onto 96-well plates. And the biotin conjugated anti- BMP-4 polyclonal antibody was used as detection antibodies. The standards, test samples and biotin conjugated detection antibody were added to the wells subsequently, and wash with wash buffer. Avidin-Biotin-Peroxidase Complex was added and unbound conjugates were washed away with wash buffer. TMB substrates were 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 BMP-4 amount of sample captured in plate. Read the O.D. absorbance at 450nm in a microplate reader, and then the concentration of BMP-4 can be calculated.

Range: 62.5 pg/ml-4000 pg/ml

Sensitivity < 4 pg/ml

Storage and Expiration: Store at 2-8℃ for 4 months, or at -20℃ for 8 months.

Application: For quantitative detection of BMP-4 in human serum, plasma, body fluids, tissue lysates or cell culture supernates.

Kit components
1.     One 96-well plate pre-coated with anti-human BMP-4 antibody
2.     Lyophilized BMP-4 standards: 2 tubes (10 ng / tube)                    
3.     Sample / Standard diluent buffer: 30ml
4.     Biotin conjugated anti-Human BMP-4 antibody (Concentrated): 130 μl. Dilution: 1:100
5.     Antibody diluent buffer: 12 ml
6.     Avidin-Biotin-Peroxidase Complex (ABC) (Concentrated): 130 μl. Dilution: 1:100
7.     ABC diluent buffer: 12 ml
8.     TMB substrate: 10 ml
9.     Stop solution: 10 ml
10.   Wash buffer (25X): 30 ml
Note: Reconstitute standards and test samples with Kit Component 3.

Gene Information

OMIM: 112262
Swiss Port: P12644
Gene ID: 652

Other Information

Please check datasheet for protocol