Description

Nori Equine MMP13 ELISA Kit Summary

Alternative names for MMP13: Matrix metalloproteinase 13

Alternative names for equine: horse

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 13 (MMP-13) also named collagenase 3 is an enzyme that in Monkeys is encoded by the MMP13 gene.[1][2] and is a member of the MMP family. Like most MMPs, it is secreted as an inactive pro-form. It is activated once the pro-domain is cleaved, leaving an active enzyme composed of the catalytic domain and the hemopexin-like domain PDB: 1PEX. Although the actual mechanism has not been described, the hemopexin domain participates in collagen degradation, the catalytic domain alone being particularly inefficient in collagen degradation. During embryonic development, MMP13 is expressed in the skeleton as required for restructuring the collagen matrix for bone mineralization. In pathological situations it is highly overexpressed; this occurs in Monkey carcinomas, rheumatoid arthritis and osteoarthritis.[3] 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 MMPs are secreted as inactive proproteins which are activated when cleaved by extracellular proteinases. The protein encoded by this gene cleaves type II collagen more efficiently than types I and III. It may be involved in articular cartilage turnover and cartilage pathophysiology associated with osteoarthritis. MMP13 expression is regulated by IL-1 receptor antagnist signalling [4] and its overexpression of MMP13 is associated with the SMAD-independent TGF-beta signaling pathway.[5] MMP13 in involved in cancer development and overexpression promotes cancer growth.[6] RKIP blocks transcriptional activation of MMP13 to inhibit local breast cancer invasion.[7]

References

1. Mitchell PG, et al. (1996) J. Clin. Invest. 97 (3), 761-768.
2. Freije JM, et al. (1994) J. Biol. Chem. 269 (24), 16766-16773 (1994)
3. Johansson N, Ahonen M, Kähäri VM (2000). Cell Mol Life Sci. 57 (1): 5–15.
4. Goto H, et al. (2015) PLoS ONE 10 (10), E0140942.
5. Aref-Eshghi E, et al. (2015) Arthritis Res. Ther. 17, 264.
6. You Y, et al. (2015) Cancer Sci. 106 (12), 1669-1677.
7. Datar I, et al. (2015) PLoS ONE 10 (8), E0134494.

Product Citations