Human MMP-3 enzyme-linked immunoassay kit
| Specification | 96 Test |
|---|---|
| Sensitivity | 0.79 ng/ml (50 μl)；4.66 ng/ml (10 μl)； |
| Standard Curve Range | 41.15~30000 pg/ml |
| Standard Curve Gradient | 7 Points/3 Folds |
| Number of Incubations | 2 |
| Detectable sample | Liquid phase sample of soluble substances. For example: serum, plasma, cell culture supernatant, tissue grinding liquid, etc. |
| Sample Volume | 50 μl/10 μl |
| Type | Fully Ready-to-Use |
| Operation Duration | 120min |
| pg/ml | O.D. | Average | Corrected | |
|---|---|---|---|---|
| 0.00 | 0.0080 | 0.0084 | 0.0082 | |
| 41.15 | 0.0319 | 0.0354 | 0.0337 | 0.0255 |
| 123.46 | 0.0831 | 0.0823 | 0.0827 | 0.0745 |
| 370.37 | 0.2282 | 0.2572 | 0.2427 | 0.2345 |
| 1111.11 | 0.6496 | 0.6786 | 0.6641 | 0.6559 |
| 3333.33 | 1.5680 | 1.6220 | 1.5950 | 1.5868 |
| 10000.00 | 2.9370 | 3.0260 | 2.9815 | 2.9733 |
| 30000.00 | 3.9709 | 4.0157 | 3.9933 | 3.9851 |
Precision
| Intra-assay Precision | Inter-assay Precision | |||||
| Sample Number | S1 | S2 | S3 | S1 | S2 | S3 |
| 22 | 22 | 22 | 6 | 6 | 6 | |
| Average(pg/ml) | 653.3 | 3162.9 | 11055.8 | 601.2 | 3117.6 | 9789.5 |
| Standard Deviation | 42.4 | 205.0 | 754.4 | 23.4 | 196.2 | 309.7 |
| Coefficient of Variation(%) | 6.5 | 6.5 | 6.8 | 3.9 | 6.3 | 3.2 |
Intra-assay Precision (Precision within an assay) Three samples of known concentration were tested twenty times on one plate to assess intra-assay precision.
Inter-assay Precision (Precision between assays) Three samples of known concentration were tested six times on one plate to assess intra-assay precision.
Spike Recovery
The spike recovery was evaluated by spiking 3 levels of human MMP-3 into health human serum sample. The un-spiked serum was used as blank in this experiment.
The recovery ranged from 80% to 107% with an overall mean recovery of 92%.
Sample Values
| Sample Matrix | Sample Evaluated | Range (ng/ml) | Detectable (%) | Mean of Detectable (ng/ml) |
|---|---|---|---|---|
| Serum | 30 | 1.47-24.25 | 100 | 9.95 |
Serum/Plasma – Thirty samples from apparently healthy volunteers were evaluated for the presence of MMP-3 in this assay. No medical histories were available for the donors.
Product Data Sheet
Background: MMP-3
Matrix metalloproteinases (MMPs), also called matrixins, constitute a family of zinc and calcium dependent endopeptidases that function in the breakdown of extracellular matrix (ECM). They play an important role in many normal physiological processes such as embryonic development, morphogenesis, reproduction and tissue remodeling. They also participate in many pathological processes such as arthritis, cancer and cardiovascular disease. While the amounts of newly synthesized MMPs are regulated mainly at the levels of transcription, the proteolytic activities of existing MMPs are controlled through both the activation of proenzymes or zymogens and the inhibition of active enzymes by endogenous inhibitors, alpha -macroglobulins and tissue inhibitors of metalloproteinases (TIMPs).
MMP-3 (also referred to as stromelysin-1) may be expressed in fibroblasts, chondrocytes, endothelial cells, macrophages, vascular smooth muscle cells, osteoblasts, and keratinocytes in response to appropriate stimuli. Various agents regulate its biosynthesis. Inflammatory cytokines such as IL-1 and TNF-alpha, epidermal growth factor, platelet-derived growth factor, phorbol and oncogenic cellular transformation are the inductive agents. In comparison, retinoic acid, glucocorticoids, estrogen, progesterone and TGF-beta suppress MMP-3 synthesis.
MMP-3 is secreted from the cells as a proenzyme. The proenzyme has been shown to stimulate plasminogen activation. The N-terminal pro-domain contains the cysteine switch motif conserved in MMPs that maintains MMP-3 in the latent state. Activation of the proenzyme results in the removal of the pro-domain. MMP-3 activation can be achieved in vitro by proteases such as itself, chyrotrypsin, neutrophil elastase and plasma kallikrein, and by mercury compounds. The resulting active enzyme consists of a catalytic domain with a zinc-binding motif conserved in metzincins. A short hinge peptide links the catalytic domain to the C-terminal hemopexin-like domain. The active MMP-3 is capable of cleaving types III, IV, IX and X collagen, aggrecan, fibronectin, laminin, IGFBP-3, serpins, and IL-1 beta. The active enzyme also activates proMMP-1, -8, -9, and -13. Therefore, it is suggested that MMP-3 may participate in physiological matrix turnover and pathological destruction of the tissue. For example, MMP-3 is required for the generation of a macrophage chemoattractant in a model of herniated disc resorption.
