Human OPG ELISA Kit

Catalog No.: BEK1179

Size: 96T

Range: 93.8 pg/ml-6,000 pg/ml

Sensitivity < 5 pg/ml

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

Application: For quantitative detection of OPG in Human serum, body fluids, tissue lysates or cell culture supernates. 

Introduction

Osteoprotegerin (OPG), also known as osteoclastogenesis inhibitory factor (OCIF), or tumor necrosis factor receptor superfamily member 11B (TNFRSF11B), is a protein that in humans is encoded by the TNFRSF11B gene. It is a member of the tumor necrosis factor (TNF) receptor superfamily, and comprising 401 amino acid residues arranged into 7 structural domains. It is a decoy receptor for the receptor activator of nuclear factor kappa B ligand (RANKL). OPG can reduce the production of osteoclasts by inhibiting the differentiation of osteoclast precursors. Elevated OPG levels has been reported in heart diseases, in Placebo effect serum responses in IBS patients and in severe mental disorders.

Principle of the Assay 

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

Kit components

1. One 96-well plate pre-coated with anti-Human OPG antibody
2. Lyophilized Human OPG standards: 2 tubes (10 ng / tube)
3. Sample / Standard diluent buffer: 30ml
4. Biotin conjugated anti-Human OPG 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