The Immunogen: Matrix metalloproteinase-2 (MMP2) (gelatinase A, 72kDa gelatinase, 72kDa type IV collagenase), a member of the matrix metalloproteinase (MMP) enzyme family, is 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. MMP2 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. Mutations in this gene have been associated with Winchester syndrome, Torg syndrome and nodulosis-arthropathy-osteolysis syndrome. MMP2 is secreted as an inactive proenzyme/zymogen which is activated when cleaved by extracellular proteinases. The proform has a molecular weight of 72kDa and the active form 62kDa. A C-terminus peptide (aa. 475-490) was selected which is highly conserved and unique to MMP2, and is present in both pro- and active forms. The peptide was conjugated to KLH and use to immunize rabbits. The hyperimmune serum was affinity purified on a peptide-coupled sepharose column. The purified immunoglobulin recognizes 72 kDa and 66 kDa bands corresponding to MMP2 on Western immunoblots. Some tissue extracts also show MMP2 complexes and degradation products. Polyclonal MMP2/475 antibody immunostains MMP2 specifically in various tissue preparations including paraffin sections. This antibody has been marketed for many years by other companies and is now produced by EnCor Biotechnology Inc. The Human Genome Organization (HUGO) Gene Nomenclature Committee (HGNC) name for this protein is simply MMP2.

Left: Blots of recombinant human MMP2 both pro and active forms (left lane) and crude homogenate from injured rat peripheral nerve (right lane) immunolabeled with RPCA-MMP2 antibody. Right: Cross-section of injured rat nerve showing MMP2 expression by Schwann cells and other reactive cells.

Antibody Characteristics: The antibody was raised against a peptide corresponding to the C terminus of MMP2, specifically MGPLLVATFWPELPEK, which was coupled to keyhole limpet hemocyanin with glutaraldehyde as described in reference 1. The antibody is provided in the form of affinity of purified IgG at a concentration of 1mg/ml. Store at 4°C or -20°C. Avoid repeat freezing and thawing.

Suggestions for use: The antibody can be diluted to 1:500-1,000 for immunofluorescence staining and 1:10,000 for western blotting. On western blots look for a major band at 66 and 72kDa, corresponding to the proenzyme (72kDa) and the activated form (66kDa).

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References:

1. Collier, I. E., Wilhelm, S. M., Eisen, A. Z., Marmer, B. L., Grant, G. A., Seltzer, J. L., Kronberger, A., He, C. S., Bauer, E. A., and Goldberg, G. I. H-ras oncogene-transformed human bronchial epithelial cells (TBE-1) secrete a single metalloprotease capable of degrading basement membrane collagen. J. Biol. Chem. 263:6579-6587 (1988).

2. C. Krekoski, D. Neubauer, J.B. Graham D. Muir. Metalloproteinase-dependent predegeneration in vitro enhances axonal regeneration within acellular peripheral nerve grafts. J. Neuroscience 22:10408-10415 (2002).

3. D. Muir. Differences in proliferation and invasion by normal, transformed and NF1 Schwann cell cultures are influenced by matrix metalloproteinase expression. Clinical and Experimental Metastasis 13:303-314 (1995).

Price and Availability: - We currently supply 100 microliter of 1mg/ml of affinity purified specific antibody for $200. Material is in stock and ready for immediate shipping.

Limitations: This product is for research use only and is not approved for use in humans or in clinical diagnosis.

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