Recommended Products
Assay
≥95% (SDS-PAGE)
Quality Level
form
lyophilized
manufacturer/tradename
Calbiochem®
storage condition
OK to freeze
avoid repeated freeze/thaw cycles
solubility
water: soluble
shipped in
wet ice
storage temp.
−70°C
General description
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.
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.
Application
Immunoblotting (see comments)
Substrate Cleavage Assay (see comments)
Zymography (see comments)
Substrate Cleavage Assay (see comments)
Zymography (see comments)
Warning
Toxicity: Standard Handling (A)
Physical form
Lyophilized from 100 mM NaCl, 50 mM HEPES, pH 7.3.
Reconstitution
Following reconstitution, aliquot into siliconized vials and freeze (-70°C).
Analysis Note
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.
Other Notes
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.
Legal Information
CALBIOCHEM is a registered trademark of Merck KGaA, Darmstadt, Germany
Storage Class Code
11 - Combustible Solids
WGK
WGK 1
Flash Point(F)
Not applicable
Flash Point(C)
Not applicable
Certificates of Analysis (COA)
Search for Certificates of Analysis (COA) by entering the products Lot/Batch Number. Lot and Batch Numbers can be found on a product’s label following the words ‘Lot’ or ‘Batch’.
Already Own This Product?
Find documentation for the products that you have recently purchased in the Document Library.
S Netzel-Arnett et al.
Analytical biochemistry, 195(1), 86-92 (1991-05-15)
Four new fluorogenic heptapeptide substrates have been synthesized with sequences that are optimized for five human matrix metalloproteinases (MMP). All four substrates are similar to one recently reported by Stack and Gray (1989, J. Biol. Chem. 264, 4277-4281) and have
D E Kleiner et al.
Analytical biochemistry, 218(2), 325-329 (1994-05-01)
Zymography is an electrophoretic technique used to identify proteolytic activity in enzymes separated in polyacrylamide gels under nonreducing conditions. It has been used extensively in the qualitative evaluation of proteases present in tumors and cell culture conditioned media. Using commercially
G Galazka et al.
Biochemistry, 35(34), 11221-11227 (1996-08-27)
Matrix metalloproteinases (MMPs) can be activated in vitro by multiple mechanisms such as treatment with proteases, organomercurials, oxidants, and detergents. The proposed cysteine switch model for activation suggests that these multiple methods for activation cause the dissociation of the single
D Cottam et al.
International journal of oncology, 2(6), 861-872 (1993-06-01)
In order for tumor cells to colonise secondary organs and tissues it is necessary for them to be able to complete all the essential steps of the metastatic cascade. We discuss here some of the important aspects of this process
Electrophoretic analysis of plasminogen activators in polyacrylamide gels containing sodium dodecyl sulfate and copolymerized substrates.
C Heussen et al.
Analytical biochemistry, 102(1), 196-202 (1980-02-01)
Our team of scientists has experience in all areas of research including Life Science, Material Science, Chemical Synthesis, Chromatography, Analytical and many others.
Contact Technical Service