Description
Introduction: Collagenase 3 (MMP13) is a secreted 452 residue protein which is released from cells as aninactive zymogen and activated extracellularly by removal of its propeptide. MMP13 was first identified in tumor cells but has since been found in synoviocytes and normal fibroblasts stimulated by IL6 or a combination of TNF alpha and IL1. Under normal conditions, MMP13 is expressed during embryogenesis (fetal bone development) and is present only at very low levels in adult tissue. However, this enzyme is reported to be involved in the development and metastasis of breast and lung carcinomas, chondrosarcomas and osteosarcomas, head and neck carcinomas and some forms of skin cancer. Additionally, this enzyme plays an important role in degenerative bone diseases including osteoarthritis and rheumatoid arthritis. In rodents, MMP13 is thought to fill the role of MMP1, which they appear to lack.
Principle of the Assay: The microtiter plate provided in this kit has been pre-coated with an antibody specific to MMP-13. Standards or samples are then added to the appropriate microtiter plate wells with a biotin-conjugated antibody preparation specific for MMP-13 and Avidin conjugated to Horseradish Peroxidase (HRP) is added to each microplate well and incubated. Then a TMB (3,3',5,5' tetramethyl-benzidine) substrate solution is added to each well. Only those wells that contain MMP-13, biotin-conjugated antibody and enzyme-conjugated Avidin will exhibit a change in color. The enzyme-substrate reaction is terminated by the addition of a sulphuric acid solution and the color change is measured spectrophotometrically at a wavelength of 450 nm +/- 2 nm. The concentration of MMP-13 in the samples is then determined by comparing the O.D. of the samples to the standard curve