Human MMP13 ELISA Kit

Catalog No.: BEK1164

Size: 96T

Range: 156pg/ml-10,000pg/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 MMP13 in human serum, plasma, body fluids, tissue lysates or cell culture supernatants.

Introduction

Matrix metallopeptidase 13 (MMP13) is a member of the matrix metalloproteinase (MMP) family. Like most MMPs it secreated as an inactive pro-form. During embryonic development, MMP13 is expressed in the skeleton as required for restructuring the collagen matrix for bone mineralization. In pathological situations it is highly overexpressed, this occurs in human carcinomas and in rheumatoid arthritis and osteoarthritis. the expression of MMP13 in osteoarthritic cartilage and its activity against type II collagen indicates that the enzyme plays a significant role in cartilage collagen degradation and must, therefore, form part of a complex target for proposed therapeutic interventions based on collagenase inhibition.

Principle of the Assay  

This kit was based on sandwich enzyme-linked immune-sorbent assay technology. Anti-MMP13 polyclonal antibody was pre-coated onto 96-well plates. And the biotin conjugated anti-MMP13 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 MMP13 amount of sample captured in plate. Read the O.D. absorbance at 450nm in a microplate reader, and then the concentration of MMP13 can be calculated.

Kit components

1. One 96-well plate pre-coated with anti-human MMP13 antibody
2. Lyophilized human MMP13 standards: 2 tubes (10 ng / tube)                     
3. Sample / Standard diluent buffer: 30ml
4. Biotin conjugated anti-human MMP13 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