Human BMP-7 ELISA Kit

Catalog No.: BEK1019

Size: 96T

Range: 31.2 pg/ml-2000 pg/ml

Sensitivity < 10pg/ml

Storage and Expiration: Store at 2-8℃ for 6 months, or at -20℃ for 12 months. 

Application: For quantitative detection of BMP-7 in human serum, plasma, urine, bone tissue or cell culture supernatants.

Introduction

Bone morphogenetic protein 7 or BMP7 (also known as osteogenic protein-1 or OP-1) is a member of the TGF-β superfamily. It is expressed in the brain, kidneys and bladder. Like other members of the bone morphogenetic protein family of proteins, it plays a key role in the transformation of mesenchymal cells into bone and cartilage. It is inhibited by noggin and a similar protein, chordin, which are expressed in the Spemann-Mangold Organizer. BMP7 induces the phosphorylation of SMAD1 and SMAD5, which in turn induce transcription of numerous osteogenic genes. It has been demonstrated that BMP7 treatment is sufficient to induce all of the genetic markers of osteoblast differentiation in many cell types.

Principle of the Assay  

This kit was based on sandwich enzyme-linked immune-sorbent assay technology. Anti-BMP-7 polyclonal antibody was pre-coated onto 96-well plates. And the biotin conjugated anti-BMP-7 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-7 amount of sample captured in plate. Read the O.D. absorbance at 450nm in a microplate reader, and then the concentration of BMP-7 can be calculated.

Kit components

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

Note: Reconstitute standards and test samples with Kit Component 3.

Material Required But Not Provided

1. 37℃ incubator
2. Microplate reader (wavelength: 450nm)
3. Precise pipette and disposable pipette tips
4. Automated plate washer
5. ELISA shaker
6. 1.5ml of Eppendorf tubes
7. Plate cover
8. Absorbent filter papers
9. Plastic or glass container with volume of above 1L