Description
Enzyme immunoassay for the quantitative determination of matrix metalloproteinase 2 (MMP-2) 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 (1). MMPs are presented in various types of cells, including cardiomyocytes, cerebral neurons, hepatocytes, and many others, thus influencing various processes.
MMP-2, also called gelatinase A, is reported to play a crucial intracellular role where it cleaves several sarcomeric proteins inside cardiac myocytes during oxidative stress-induced injury (2). Elevated levels of MMP-2 (combined with MMP-9) are also associated with highly tumorigenic cancers, such as breast and gastric cancers.
Other Names
Matrix metalloproteinase-2 (MMP-2), MMP2, 72 kDa type IV collagenase, 72 kDa gelatinase, Gelatinase A, TBE-1, UniProtKB# P08253, EC:3.4.24.24.
Principle of Method
The microtiter plate is coated with the antibody specifically binding the Matrix metalloproteinase 2. 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-2 in the specimen. The concentration of MMP-2 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: 0.625 ng/mL - 40 ng/mL
Sensitivity
The limit of detection, defined as a concentration of human MMP-2 giving absorbance higher than absorbance of blank + 3 standard deviations, is better than 0.25 ng/mL of sample.
Precision
Intra-Assay
| Test sample | Mean (ng/mL) | SD | CV |
| 1 |
175 |
5.1 |
3% |
| 2 |
208 |
4.3 |
2% |
Inter-Assay (Run-to-Run)
| Test sample | Mean (ng/mL) | SD | CV |
| 1 |
252 |
21.7 |
9% |
| 2 |
145 |
4.3 |
3% |
Accuracy
Dilution Linearity:
| Test sample | Dilution |
Measured Concentration (ng/mL) |
Expected Concentration (ng/mL) |
Yield |
| 1 |
175 |
- | - | |
| 1 | 2x | 96 |
88 |
110% |
| 1 | 4x | 50 |
44 |
114% |
| 1 | 8x | 21 |
22 |
97% |
| 2 |
150 |
- |
- | |
| 2 | 2x | 80 |
75 |
107% |
| 2 | 4x | 43 |
37 |
114% |
| 2 | 8x | 19 |
19 |
99% |
Spiking Recovery:
| Test sample | Spike (ng/mL) |
Measured Concentration (ng/mL) |
Expected Concentration (ng/mL) |
Yield |
| 1 |
- | 208 |
- | - |
| 1 | 250 |
453 |
458 |
99% |
| 1 | 125 |
334 |
333 |
100% |
| 1 | 62.5 |
270 |
270 |
100% |
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
- Łukaszewicz-Zając M, Mroczko B, Słowik A. Matrix metalloproteinases (MMPs) and their tissue inhibitors (TIMPs) in amyotrophic lateral sclerosis (ALS). J Neural Transm (Vienna). 2014 Nov;121(11):1387-97. doi: 10.1007/s00702-014- 1205-3. Epub 2014 Jul 22. PMID: 25047909; PMCID: PMC4210652.
- Bassiouni W, Ali MAM, Schulz R. Multifunctional intracellular matrix metalloproteinases: implications in disease. FEBS J. 2021 Dec;288(24):7162-7182. doi: 10.1111/febs.15701. Epub 2021 Jan 22. PMID: 33405316.
Related Products
- QuickZyme Human MMP-2 Activity Assay (measures active & total MMP-2)
-
Matrix Metalloproteinase Assays
- Extracellular Matrix Assays
- Immunoassays
Ilex Life Sciences LLC is an authorized distributor of PromedeusLab.