Nori Porcine MMP14 ELISA Kit
Price range: $508.00 through $916.00
This ELISA kit is for quantification of MMP14 in porcine. 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 MMP14 has been pre-coated onto a microplate. Standards and samples are pipetted into the wells and any MMP14 present is bound by the immobilized antibody. After washing away any unbound substances, a detection antibody specific for MMP14 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 MMP14 bound in the initial step. The color development is stopped, and the intensity of the color is measured.
Alternative names for MMP14: Matrix metalloproteinase-14, MMP-14
This product is for Laboratory Research Use Only not for diagnostic and therapeutic purposes or any other purposes.
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Description
Nori Porcine MMP14 ELISA Kit Summary
Alternative names for MMP14: Matrix metalloproteinase-14, MMP-14,
Alternative names for porcine: Pig, swine
| 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 | Q9XT90 |
| 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 | 60 pg/mL |
| Detection Range | 0.313-20 ng/mL |
| Specificity | Porcine MMP14 |
| 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-14 (MMP14) is an enzyme that is encoded by the MMP14 gene.[1] MMP14 belongs to the matrix metalloproteinase (MMP) family that 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 pro-proteins which are activated when cleaved by extracellular proteinases. However, MMP14 is a member of the membrane-type MMP (MT-MMP) subfamily; each member of this subfamily contains a potential transmembrane domain suggesting that these proteins are expressed at the cell surface rather than secreted. MMP14 has been shown to interact with TIMP2.[2] MMP14 plays an important role in extracellular matrix by degrading type I collagen, activating pro-MMP2, by processing cell adhesion molecules such as CD44 and integrin alpha.[3] MMP14 is therefore a key enzyme in many physiological and pathological processes such as angiogenesis and tumor invasion.
References
- Sato H, et al. (1994). Nature 370 (6484): 61–5.
- Zucker S, et al. (1998). J. Biol. Chem. 273 (2): 1216–22.
- Seike M (2003) Cancer Lett 194:1.
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