Nori Canine MMP-2 ELISA Kit
$461.00 – $832.00
This ELISA kit is for quantification of MMP-2 in dog. This is a quick ELISA assay that reduces time to 50% compared to the conventional method, and the entire assay only takes 3 hours. This assay employs the quantitative sandwich enzyme immunoassay technique and uses biotin-streptavidin chemistry to improve the performance of the assays. An antibody specific for MMP-2 has been pre-coated onto a microplate. Standards and samples are pipetted into the wells and any MMP-2 present is bound by the immobilized antibody. After washing away any unbound substances, a detection antibody specific for MMP-2 is added to the wells. Following wash to remove any unbound antibody reagent, a detection reagent is added. After intensive wash a substrate solution is added to the wells and color develops in proportion to the amount of MMP-2 bound in the initial step. The color development is stopped, and the intensity of the color is measured.
Alternative names for MMP-2: Matrix metalloproteinase-2 (MMP-2), 72 kDa type IV collagenase, gelatinase A
This product is for Laboratory Research Use Only not for diagnostic and therapeutic purposes or any other purposes.
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Description
Nori Canine MMP-2 ELISA Kit Summary
Alternative names for MMP-2: Matrix metalloproteinase-2 (MMP-2), 72 kDa type IV collagenase, gelatinase A
Alternative name for canine: Dog
| Assay Type | Solid Phase Sandwich ELISA |
| Format | 96-well Microplate or 96-Well Strip Microplate |
| Method of Detection | Colorimetric |
| Number of Targets Detected | 1 |
| Target Antigen Accession Number | A0A8I3MGQ5 |
| Assay Length | 3 hours |
| Quantitative/Semiquantitative | Quantitative |
| Sample Type | Plasma, Serum, Cell Culture, Urine, Cell/Tissue Lysates, Synovial Fluid, BAL, |
| Recommended Sample Dilution (Plasma/Serum) | No dilution for sample <ULOQ; sufficient dilution for samples >ULOQ |
| Sensitivity | 50 pg/mL |
| Detection Range | 250-16000 pg/mL |
| Specificity | Natural and recombinant canine MMP-2 |
| Cross-Reactivity | < 0.5% cross-reactivity observed with available related molecules, < 50% cross-species reactivity observed with species tested. |
| Interference | No significant interference observed with available related molecules |
| Storage/Stability | 4 ºC for up to 6 months |
| Usage | For Laboratory Research Use Only. Not for diagnostic or therapeutic use. |
| Additional Notes | The kit allows for use in multiple experiments. |
Standard Curve
Kit Components
1. Pre-coated 96-well Microplate
2. Biotinylated Detection Antibody
3. Streptavidin-HRP Conjugate
4. Lyophilized Standards
5. TMB One-Step Substrate
6. Stop Solution
7. 20 x PBS
8. Assay Buffer
Other Materials Required but not Provided:
1. Microplate Reader capable of measuring absorption at 450 nm
2. Log-log graph paper or computer and software for ELISA data analysis
3. Precision pipettes (1-1000 µl)
4. Multi-channel pipettes (300 µl)
5. Distilled or deionized water
Protocol Outline
1. Prepare all reagents, samples and standards as instructed in the datasheet.
2. Add 100 µl of Standard or samples to each well and incubate 1 h at RT.
3. Add 100 µl of Working Detection Antibody to each well and incubate 1 h at RT.
4. Add 100 µl of Working Streptavidin-HRP to each well and incubate 20 min at RT.
5. Add 100 µl of Substrate to each well and incubate 5-30 min at RT.
6. Add 50 µl of Stop Solution to each well and read at 450 nm immediately.
Background:
Matrix metalloproteinase-2 (MMP-2) is known as 72 kDa type IV collagenase, and gelatinase A is an enzyme that in humans is encoded by the MMP2 gene.(1) Proteins of the MMP family are involved in the breakdown of extracellular matrix in normal physiological processes, such as embryonic development, reproduction, and tissue remodeling, as well as in disease processes, such as arthritis and metastasis. Most MMP’s are secreted as inactive proproteins which are activated when cleaved by extracellular proteinases. This gene encodes an enzyme which degrades type IV collagen, the major structural component of basement membranes. The enzyme plays a role in endometrial menstrual breakdown, regulation of vascularization and the inflammatory response. Activation of MMP-2 requires a proteolytic processing. A complex of membrane type 1 MMP (MT1-MMP/MMP14) and tissue inhibitor of metalloproteinase 2 recruits pro-MMP 2 from extracellular milie to the cell surface. Activation then requires active molecule of MT1-MMP and auto catalytic cleavage. Clustering of integrin chain promotes activation of MMP-2. Another factor that will support the activation of MMP-2 is cell-cell clustering. A wild-type activated leukocyte cell adhesion molecule (ALCAM) also required to activate MMP-2. Mutations in the MMP2 gene are associated with Torg-Winchester syndrome, multicentric osteolysis and arthritis syndrome.(2) MMP2 has been shown to interact with THBS2,(3) TIMP2,(4-7) Thrombospondin 1,(4) CCL7 (8) and TIMP4.(6, 7)
Reference
- Devarajan P, et al. Biol. Chem. 267 (35): 25228–32.
- Martignetti JA, et al. Genet. 28 (3): 261–5.
- Bein K, Simons M. Biol. Chem. 275 (41): 32167–73.
- Morgunova E, et al. Natl. Acad. Sci. U.S.A. 99 (11): 7414–9.
- Overall CM, et al. Biol. Chem. 275 (50): 39497–506.
- Bigg HF, et al. Biol. Chem. 272 (24): 15496–500.
- Kai HS, et al. Biol. Chem. 277 (50): 48696–707.
- McQuibban GA, et al. Science 289 (5482): 1202–6.
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