AffinityImmuno Products

Introduction

MMP9 encodes gelatinase B, a secreted zinc-dependent enzyme released by neutrophils, macrophages and several other cell types. Once activated by cleavage of its inactive proform, the enzyme breaks down type IV and V collagen along with fibronectin, allowing it to remodel the extracellular matrix. This remodeling activity supports normal processes such as embryonic development, wound repair and the mobilization of bone-marrow progenitor cells, while excessive or dysregulated activity contributes to inflammatory joint disease, tumor invasion and metastatic spread. Given its dual role in tissue homeostasis and pathological remodeling, MMP9 is widely measured as a marker of inflammation and matrix turnover.

MMP9’s dual role in normal tissue remodeling and pathological processes makes it one of the most frequently measured matrix metalloproteinases in research spanning inflammatory joint disease, wound healing, and cancer invasion/metastasis studies, often alongside other MMP-family members and their endogenous inhibitors (TIMPs) in multi-marker panels.

Principle of the Assay

This kit utilizes a sandwich enzyme immunoassay format. The microtiter plate is pre-coated with a capture antibody specific to Human MMP9; a separate set of standard plate wells pre-coated by protein-related techniques is provided for the standard curve. Standards or samples are dispensed into the appropriate wells, followed by addition of an HRP-conjugated detection antibody that also targets Human MMP9. Upon addition of TMB substrate solution, a colorimetric signal develops exclusively in wells containing both Human MMP9 and the HRP-conjugated antibody. The enzymatic reaction is terminated by addition of sulfuric acid stop solution, and absorbance is measured at 450 nm (± 10 nm). Sample concentrations are then determined by comparing the optical density of each sample against the generated standard curve.