PF063

MMP-3, Proenzyme, Human, Recombinant

• Comisión internacional de enzimas: 3.4.24.17 (BRENDA, IUBMB)
• Código UNSPSC: 12352202
• NACRES: NA.77

Propiedades

• Ensayo: ≥95% (SDS-PAGE)
• Nivel de calidad: 100
• Formulario: lyophilized
• fabricante / nombre comercial: Calbiochem^®
• condiciones de almacenamiento: OK to freeze; avoid repeated freeze/thaw cycles
• solubilidad: water: soluble
• Condiciones de envío: wet ice
• temp. de almacenamiento: −70°C

Descripción

Descripción general

Recombinant, human pro-MMP-3 purified from cell culture supernatant. May be used as a positive control or standard for zymographic analysis, or substrate assay. Requires activation for immunoblotting, prior to use. M.W. 57000/58000.

Recombinant, human pro-MMP-3 purified from cell culture supernatant. May be used as a positive control or standard for zymographic analysis, or substrate assay. Requires activation for immunoblotting, prior to use. M.W. 57000/58000.



Matrix metalloproteinases (MMPs) are a family of enzymes that are responsible for the degradation of extracellular matrix components such as collagen, laminin and proteoglycans. In addition to sequence homology, all MMPs share the following characteristics: the catalytic mechanism is dependent upon a zinc ion at the active center, they cleave one or more extracellular matrix components, they are secreted as zymogens which are activated by removal of an approximately 10 kDa segment from the N terminus and they are inhibited by tissue inhibitor of metalloproteinases (TIMP). These enzymes are involved in normal physiological processes such as embryogenesis and tissue remodeling and may play an important role in angiogenesis, arthritis, periodontitis, and metastasis. Matrix metalloproteinase-3 (MMP-3) also known as stromelysin-1 and transin (EC 3.4.24.17) cleaves a number of substrates including cartilage proteoglycan, collagen types II, III, IV, V and IX, fibronectin, laminin, and can activate MMP 1. MMP-3 is secreted as ~57 and ~59 kDa proenzymes and can be activated in vitro by organomercurials (e.g., 4 aminophenylmercuric acetate, APMA) and in vivo by proteases via intermediate forms to a 45 kDa active MMP 3 enzyme. Further autolysis to a ~28 kDa form can also occur. MMP-3 is thought to play an important role in pathophysiological degradation processes associated with conditions such as rheumatoid arthritis and cancer cell invasion.

Aplicación

Immunoblotting (see comments)

Substrate Cleavage Assay (see comments)

Zymography (see comments)

Advertencia

Toxicity: Standard Handling (A)

Forma física

Lyophilized from 100 mM NaCl, 50 mM HEPES, pH 7.3.

Reconstitución

Following reconstitution, aliquot into siliconized vials and freeze (-70°C).

Nota de análisis

The activity of proenzyme MMP 3 was measured by substrate cleavage assay using 0.5 mM thiopeptiolide (Ac-Pro-Leu-Gly-S-Leu-Leu-Gly-Oet) as a substrate. The activity was also assessed by degradation of a peptide substrate (DNP-PYAYWMR) using activated MMP-3 as measured by HPLC.

Otras notas

Proenzyme MMP-3 may be used as a positive control or standard for immunoblotting, zymographic analysis, or substrate cleavage assays. 0.5 μg/lane was used for SDS-PAGE and immunoblotting. For zymography with casein 1μg/lane of Proenzyme MMM-3 or activated MMP-3 was used. Proenzyme MMP-3 can be activated in vitro by incubation in 50 mM Tris, pH 7.5, containing 0.05% Triton-X-100, 5 mM CaCl₂ and 1 mM 4 aminophenyl mercuric acetate (APMA) for 2-4 hours at 37°C. To dissolve APMA, make a 10 mM stock solution in 0.05 M NaOH. Approximately 90% of proenzyme MMP-3 is activated with a 4 hour incubation at 37°C using 1 mM APMA.

Información legal

CALBIOCHEM is a registered trademark of Merck KGaA, Darmstadt, Germany

Información de seguridad

• Código de clase de almacenamiento: 11 - Combustible Solids
• Clase de riesgo para el agua (WGK): WGK 1
• Punto de inflamabilidad (°F): Not applicable
• Punto de inflamabilidad (°C): Not applicable