• Product NameHuman MMP-9 ELISA kit
• Brief DescriptionELISA Kit
• ApplicationsELISA
• Species ReactivityHu
• SpecificityNatural and recombinant Human MMP-9 Ligand
• Crossing ReactivityNo significant interference observed with available related molecules.
• Target NameHuman MMP-9

• Detect Range: 62.5-4000pg/ml
  Sensitivity: 31pg/mL
  Sample Type: Cell culture supernatant, serum, plasma (EDTA, citrate, heparin)
  Sample Volume: 20 uL
  Assay Time: 3 hours
  Detection method: Colorimetric

• • Aluminium pouches with a Microwell Plate coated with antibody to human MMP-9 (8x12)
  • 1 vials human MMP-9 Standard lyophilized, 4000 pg/ml upon reconstitution
  • 1 vials concentrated Biotin-Conjugate anti-human MMP-9 antibody
  • 1 vials Streptavidin-HRP solution
  • 2 bottle Standard /sample Diluent
  • 1 bottle Biotin-Conjugate antibody Diluent
  • 1 bottle Streptavidin-HRP Diluent
  • 1 bottle Wash Buffer Concentrate 20x (PBS with 1% Tween-20)
  • 1 vial Substrate Solution
  • 1 vial Stop Solution
  • 2 pieces Adhesive Films
  • package insert

Matrix metalloproteinases (MMPs) are a family of zinc-dependent endopeptidases that degrade extracellular matrix proteins (1 - 3). They are secreted as zymogens (Pro-MMPs) that are activated by a variety of proteinases or by reaction with organic mercurials. They are inhibited by specific tissue inhibitors of metalloproteinases (TIMPs) and by 2-macroglobulin (3 - 6). The regulation of MMP activity is important in tissue remodeling, inflammation, tumor growth and metastasis (3, 7 - 9).

Human MMP-9 (also known as gelatinase B) is secreted as a 92 kDa zymogen (2, 3). Cleavage of Pro-MMP-9 at or near residue 87 results in the active enzyme with a mass of approximately 82 kDa (1). MMP-9 has three fibronectin type II domains, a hemopexin-like domain and a proline-rich type V collagen-homologous domain (1 - 3). Pro-MMP-9 can be activated by MMP-3 (5) or by certain bacterial proteinases (10). MMP-9 is inhibited byα2-macroglobulin or by TIMP-1 (3 - 6), which binds to Pro-MMP-9 as well as to active MMP-9 (3). In vitro treatment of Pro-MMP-9 with 4-aminophenylmercuric acid (APMA) produces not only the 82 kDa active enzyme but also a C-terminal truncated form of approximately 65 kDa with the activity comparable to that of the 82 kDa form (11).

Pro-MMP-9 is secreted by monocytes, macrophages, neutrophils, keratinocytes, fibroblasts, osteoclasts, chondrocytes, skeletal muscle satellite cells, endothelial cells, and various tumor cells(1, 7- 21). Pro-MMP-9 expression is upregulated by TGF-β1, IL-1β, TGF-α, PDGF-AB, TNF-α, and IL-1α (7, 15, 17). Substrates for MMP-9 include denatured collagen type I (gelatin), native collagens type IV, V, VII, X and XI, fibrinogen, vitronectin, IL-1β, and entactin, a molecule that bridges laminin and type IV collagen (3, 4, 6, 13, 21 - 23).

Wilhelm, S.M. et al. (1989) J. Biol. Chem. 264:17213.

Matrisian, L.M. (1992) BioEssays 14:455.

Birkedal-Hansen, H. et al. (1993) Crit. Rev. Oral Biol. Med. 4:197.

Birkedal-Hansen, H. (1995) Curr. Opin. Cell Biol. 7:728.

Ogata, Y. et al. (1992) J. Biol. Chem. 267:3581.

Sires, U.I. et al. (1993) J. Biol. Chem. 268:2069.

Lyons, J.G. et al. (1993) J. Biol. Chem. 268:19143.

Okada, Y. et al. (1995) Lab. Invest. 72:311.

Tamura, T. et al. (1996) Endocrinology 137:3729.

Okamoto, T. et al. (1997) J. Biol. Chem. 272:6059.

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• Representative standard curve for MMP-9 ELISA. MMP-9 was diluted in serial two-fold steps in Sample Diluent.

• Flow Cytometry (FCM) General Protocol
• Immunofluorescence(IC)
• Immunohistochemistry(Polymer)
• Western Blotting