Nori Human MMP-8 ELISA Kit
$461.00 – $832.00
This ELISA kit is for quantification of MMP-8 in human. 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-8 has been pre-coated onto a microplate. Standards and samples are pipetted into the wells and any MMP-8 present is bound by the immobilized antibody. After washing away any unbound substances, a detection antibody specific for MMP-8 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-8 bound in the initial step. The color development is stopped, and the intensity of the color is measured.
Alternative names for MMP-8: Matrix metalloproteinase-8 (MMP-8), Neutrophil collagenase, PMNL collagenase (MNL-CL), CLG1, MMP8
This product is for Laboratory Research Use Only not for diagnostic and therapeutic purposes or any other purposes.
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Description
Nori Human MMP-8 ELISA Kit Summary
Alternative names for MMP-8: Matrix metalloproteinase-8 (MMP-8), Neutrophil collagenase, PMNL collagenase (MNL-CL), CLG1, MMP8
| 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 | P22894 |
| 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 | 18 pg/mL |
| Detection Range | 93.75-6000 pg/mL |
| Specificity | Human MMP-8 |
| 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:
Neutrophil collagenase, also known as matrix metalloproteinase-8 (MMP-8) or PMNL collagenase (MNL-CL), is a collagen cleaving enzyme which is present in the connective tissue of most mammals. In humans, the MMP-8 protein is encoded by the MMP8 gene.[1][2] 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. However, the enzyme encoded by this gene is stored in secondary granules within neutrophils and is activated by autolytic cleavage. Its function is degradation of type I, II and III collagens. The gene is part of a cluster of MMP genes which localize to chromosome 11q22.3. The reciprocal positive interplay between MMP-8 and TGF-beta1 contributes to HCC invasion and metastasis by inducing EMT mainly through the PI3K/Akt/Rac1 pathway[3] Study showed that miR-539 plays a key role in inhibiting osteosarcoma cell invasion and migration and can regulate MMP8 expression in osteosarcoma cells.[4] salivary levels of the analyzed biomarkers MMP-8, -9, MPO are associated with periodontal status. However, these biomarkers could not differentiate between patients with or without a MI.[5]
References
- Hasty KA, et al. (1990). J. Biol. Chem. 265(20): 11421–4.
- Devarajan P, et al. (1991). Blood. 77(12): 2731–8.
- Qin J, et al. (2016) Cancer Lett. 374 (1), 85-95.
- Jin H, et al. (2015) Int J Clin Exp Pathol 8 (7), 8075-8082.
- Rathnayake N, et al. (2015) PLoS ONE 10 (7), E0126370.
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