Description
Enzyme immunoassay for the quantitative determination of matrix metalloproteinase 3 (MMP-3) in human serum and plasma samples. Quality controls are included with the kit. Developed and manufactured in Europe by PromedeusLab.
Summary
Matrix metalloproteinases (MMPs) are zinc-dependent endopeptidases, responsible for the integrity of the basement membrane (BM) via degradation of extracellular matrix and BM components. MMPs are presented in various types of cells, including cardiomyocytes, cerebral neurons, hepatocytes, and many others, thus influencing various processes.
Matrix metalloproteinase-3 (MMP-3), also called stromelysin 1, is reported to be found in the cytosol of dopaminergic cells where its activation under stress conditions contributes to neuronal degradation (1). In chondrocytes, nuclear MMP-3 is suggested to be involved in the development of inflammatory arthritic diseases (2). MMP-3 is implicated in the formation of atherosclerotic plaques, and the MMP-3 −1612 5A/6A polymorphism is associated with myocardial infarction (MI) and stable coronary artery disease (CAD) (3).
Other Names
Matrix metalloproteinase-3 (MMP-3), MMP3, Stromelysin-1 (SL-1), Stromelysin 1 (SL1), STMY1, Transin-1, UniProtKB# P08254, EC:3.4.24.17.
Principle of Method
The microtiter plate is coated with the antibody specifically binding the Matrix metalloproteinase 3. The human serum or plasma is incubated in the plate with the capture antibody.
The specimen is washed out and the specifically bound protein is incubated with biotin-labelled detection antibody. Following another washing step, Streptavidin-HRP conjugate is added into the well. Unbound reagent is then washed out. Horseradish peroxidase (HRP) bound in the complex reacts with the chromogenic substrate (TMB) creating the blue colour. The reaction is stopped by addition of STOP solution (H2SO4).
The absorbance values are measured at 450 nm (optionally 450/630 nm) and are proportional to the concentration of MMP-3 in the specimen. The concentration of MMP-3 in unknown samples is determined from the calibration curve which is created by plotting the absorbance values against the standard concentration values.
Sample Types
Serum, Plasma
Typical Standard Curve
Assay Range: 0.2 ng/mL - 10 ng/mL
Sensitivity
The limit of detection, defined as a concentration of human MMP-3 giving absorbance higher than absorbance of blank + 3 standard deviations, is better than 0.05 ng/mL of sample.
Precision
Intra-Assay
| Test sample | Mean (ng/mL) | SD | CV |
| 1 |
16.1 |
0.4 |
2% |
| 2 |
21.8 |
1.3 |
6% |
Inter-Assay (Run-to-Run)
| Test sample | Mean (ng/mL) | SD | CV |
| 1 |
20.5 |
1.2 |
6% |
| 2 |
6.0 |
0.1 |
2% |
Accuracy
Dilution Linearity:
| Test sample | Dilution |
Measured Concentration (ng/mL) |
Expected Concentration (ng/mL) |
Yield |
| 1 |
16.0 |
- | - | |
| 1 | 2x | 8.0 |
8.0 |
100% |
| 1 | 4x | 3.7 |
4.0 |
93% |
| 1 | 8x | 1.9 |
2.0 |
96% |
| 2 |
21.1 |
- |
- | |
| 2 | 2x | 10.6 |
10.5 |
101% |
| 2 | 4x | 5.0 |
5.3 |
94% |
| 2 | 8x | 2.2 |
2.6 |
84% |
Spiking Recovery:
| Test sample | Spike (ng/mL) |
Measured Concentration (ng/mL) |
Expected Concentration (ng/mL) |
Yield |
| 1 |
- | 15.8 |
- | - |
| 1 | 30.0 |
51.4 |
45.8 |
112% |
| 1 | 15.0 |
31.0 |
30.8 |
101% |
| 1 | 7.5 |
23.7 |
23.3 |
101% |
Storage
The kit must be stored at 2–8 °C. The opened components can be stored for one week at 2–8 °C.
Usage
This assay is for in vitro research use only and is not for use in diagnostic procedures.
Resources
Summary References
- Choi, D. H., Kim, E. M., Son, H. J., Joh, T. H., Kim, Y. S., Kim, D., Flint Beal, M., & Hwang, O. (2008). A novel intracellular role of matrix metalloproteinase-3 during apoptosis of dopaminergic cells. Journal of neurochemistry, 106(1), 405–415. https://doi.org/10.1111/j.1471-4159.2008.05399.x
- Eguchi, T., Kubota, S., Kawata, K., Mukudai, Y., Uehara, J., Ohgawara, T., Ibaragi, S., Sasaki, A., Kuboki, T., & Takigawa, M. (2008). Novel transcription-factor-like function of human matrix metalloproteinase 3 regulating the CTGF/CCN2 gene. Molecular and cellular biology, 28(7), 2391–2413. https://doi.org/10.1128/MCB.01288-07
- Samnegård, A., Silveira, A., Lundman, P., Boquist, S., Odeberg, J., Hulthe, J., McPheat, W., Tornvall, P., Bergstrand, L., Ericsson, C. G., Hamsten, A., & Eriksson, P. (2005). Serum matrix metalloproteinase-3 concentration is influenced by MMP-3 -1612 5A/6A promoter genotype and associated with myocardial infarction. Journal of internal medicine, 258(5), 411–419. https://doi.org/10.1111/j.1365-2796.2005.01561.x
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Ilex Life Sciences LLC is an authorized distributor of PromedeusLab.