Description
Introduction: 72 kDa type IV collagenase also known as matrixmetalloproteinase-2 (MMP-2) and gelatinase A is an enzyme thatin humans is encoded by the MMP2 gene.Proteins of the matrix metalloproteinase (MMP) family are involvedin the breakdown of extracellular matrix in normal physiologicalprocesses, such as embryonic development, reproduction, andtissue remodeling, as well as in disease processes, such asarthritis and metastasis. Most MMP's are secreted as inactiveproproteins which are activated when cleaved by extracellularproteinases. This gene encodes an enzyme which degrades typeIV collagen, the major structural component of basementmembranes. The enzyme plays a role in endometrial menstrualbreakdown, regulation of vascularization and the inflammatoryresponse
Principle of the Assay: The microtiter plate provided in this kit has been pre-coated with an antibody specific to MMP-2. Standards or samples are then addedto the appropriate microtiter plate wells with a biotin-conjugatedantibody preparation specific for MMP-2 and Avidin conjugated toHorseradish Peroxidase (HRP) is added to each microplate welland incubated. Then a TMB (3,3',5,5' tetramethyl-benzidine)substrate solution is added to each well. Only those wells thatcontain MMP-2, biotin-conjugated antibody andenzyme-conjugated Avidin will exhibit a change in color. Theenzyme-substrate reaction is terminated by the addition of asulphuric acid solution and the color change is measuredspectrophotometrically at a wavelength of 450 nm +/- 2 nm. Theconcentration of MMP-2 in the samples is then determined bycomparing the O.D. of the samples to the standard curve