Matrix Metalloproteinase 3 (MMP-3) Activity Fluorometric Assay Kit (E-BC-F061)
For research use only.
| Detection Principle |
Matrix metalloproteinase 3 (MMP-3) is an important member of the MMP family. The precursor of MMP-3 was cleaved by plasmin, chymotrypsin and other serine proteases to remove the precursor peptide containing cysteine switch and activate MMP-3 with protease activity. MMP-3 can degrade or shear a variety of extracellular matrix components, precursor proteins or precursor enzymes, and can destroy the histological barrier of tumor cell invasion, release E-cadherin, promote tumor invasion and metastasis, and promote inflammatory response, which has received increasing attention in tumor research. In addition, MMP-3 is also involved in a series of physiological and pathological processes such as tissue morphogenesis, injury repair and inflammatory response, and plays an important role in the occurrence and development of diseases such as rheumatic arthritis and atherosclerosis. This kit is detected by fluorescence resonance energy transfer (FRET) method. MCA and Dnp are connected to two ends on the natural substrate of MMP-3 enzyme. When MMP-3 protease does not cut the substrate, the two groups are close enough to undergo fluorescence resonance energy transfer, that is, Dnp can quench the fluorescence of MCA, resulting in undetectable fluorescence. When the substrate is cut by MMP-3 protease, the ends and ends of the polypeptide are separated, the two groups are separated, the fluorescence of MCA is no longer extinguished by Dnp, and the fluorescence of MCA can be detected, so that the enzyme activity of MMP-3 protease can be detected very sensitively through fluorescence detection. MCA has a maximum excitation wavelength of 325 nm and a maximum emission wavelength of 393 nm. |
| Sample Type | Serum,Plasma,Animal tissue,Cell |
| Detection Method | Fluorometric method |
| Detection Instrument | Fluorescence microplate reader (Ex/Em=325 nm/393 nm) |
| Research Area | Amino Acids And Proteins |
| Other Reagents Required | Normal saline (0.9% NaCl) |
| Storage | This product can be stored at -20°C for 12 months with shading light. |
| Valid Period | 12 months |
| Sensitivity | 0.03 U/L |
| Detection Range | 0.03-9.66 U/L |
| Precision | inter-assay CV: 7.5-7.7% | intra-assay CV: 2.0-4.0% |
| Sample Volume | 10 μL |
| Assay Time | 50 min |
The recommended dilution factor for different samples is as follows (for reference only):
| Sample Type | Dilution Factor |
|---|---|
| 10% Rat liver tissue homogenization | 1-2 |
| 10% Rat lung tissue homogenization | 1-2 |
| 10% Mouse liver tissue homogenization | 1-2 |
| 1 × 10^6 CHO cell | 1 |
| Rat serum | 1 |
| Rat plasma | 1 |
The diluent is normal saline (0.9% NaCl) or PBS (0.01 M, pH 7.4). For the dilution of other sample types, please do pretest to confirm the dilution factor.
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