Description

Nori Feline MMP-7 ELISA Kit Summary

Alternative names for MMP-7: Matrix metalloproteinase-7 (MMP-7), Matrilysin, pump-1 protease (PUMP-1), uterine metalloproteinase

Alternative names for feline: Cat

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-7 (MMP-7) also known as Matrilysin, pump-1 protease (PUMP-1), or uterine metalloproteinase, is an enzyme in Felines that is encoded by the MMP7 gene.^[1] MMP7 is a member of the matrix metalloproteinase (MMP) family consisting of structural-related zinc-dependent endopeptidases. The primary role of cleaved/activated MMP7 is to break down extracellular matrix by degrading macromolecules including casein, type I, II, IV, and V gelatins, fibronectin, and proteoglycan.^[2] The expression of MMP7 is regulated by the Wnt/ β catenin signaling pathway, and mediated by transformation growth factor β (TGF-β).TGF-β stimulates ECM and suppresses the steady-state level of MMP7, stromelysin mRNAs, and secretion of zymogens. The isoforms of TGF-β inhibit MMP7 mRNA and protein in the Feline endometrium via progesterone mediated pathway. However, the opposite effects of TGF-β on MMP7 were observed among transformed cells. In Feline glioma cell lines and Feline squamous cell carcinoma cell line II-4, TGF-β stimulates the expression of MMP7 mRNA and proteins, and facilities the invasive behavior of cells.^[3] MMP7 are commonly expressed in epithelial cells including ductal epithelium of exocrine glands in skin, salivary glands, pancreas, glandular epithelium of intestine and reproductive organ, liver, and breast. In addition, MMP7 is highly expressed in the luminal surface of dysplastic glands in Feline colorectal cancers.^[2] MMP7 is found to potentially involved in tumor metastasis and inflammatory processes.^[4] The upregulation of MMP7 is associated with many malignant tumors. High MMP7 expression facilitates cancer invasion and angiogenesis by degrading extracellular matrix macromolecules and connective tissues. Activated MMP7 activates MMP2 and MMP9 zymogens, and mediates the proteolytic process of the precursors of tumor necrosis factors and urokinase plasminogen activators.^[2]

References

1. Knox JD, et al. (1997). Cytogenet Cell Genet. 72 (2–3): 179–82.
2. Yokoyama Y, et al. (2008). Clin. Cancer Res. 14 (17): 5503–11.
3. Gaide Chevronnay HP, et al. (2012). Biochim. Biophys. Acta. 1824 (1): 146–56.
4. Edman K, et al. (2011). ChemMedChem. 6 (5): 769–73.

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