| ALX-200-419-C005 | 5 µg | 314.00 USD |  |
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| Alternative Name: | Matrix metalloproteinase 2, Gelatinase A, 72 kDa Type IV collagenase |
| MW: | ~72kDa |
| Source: | Isolated from human rheumatoid synovial fibroblasts. Requires activation. |
| EC: | 3.4.24.24 |
| UniProt ID: | P08253 |
| Formulation: | Liquid. In 50mM TRIS-HCl, pH 7.0, containing 200mM NaCl, 5mM CaCl2, 1µM ZnCl2, 0.05% BRIJ 35 and 0.05% sodium azide. |
| Purity: | ≥90% (SDS-PAGE, Western blot) |
| Purity Detail: | No other MMP contaminants are detectable. |
| Specific Activity: | ≥850mU/mg protein (Y. Masui, et al.; Biochem. Med. 17, 215 (1977)). One unit is defined as the amount of enzyme that hydrolyzes 1µmol Dnp-Pro-Gln-Gly-Ile-Ala-Gly-Gln-D-Arg-OH per min. at 37°C, pH 7.0. |
| Application Notes: | Immunogen for antibody generation, control in immunoassays and for characterizing interactions with MMP inhibitors. |
| Shipping: | Dry Ice |
| Long Term Storage: | -80°C |
| Handling: | Avoid freeze/thaw cycles. |
| Technical Info/Product Notes: | Activity: Specific activity can be assayed with the synthetic substrate N-(2,4)-dinitrophenyl-Pro-Gln-Gly-Ile-Ala-Gly-Gln-D-Arg (Dnp-peptide) (Masui et al.). Substrate concentration should be 0.5mg/ml in 50mM TRIS-HCl, pH 7.0, 200mM NaCl, 5mM CaCl2, 1µM ZnCl2, 0.05% sodium azide, 0.05% BRIJ35, containing 0.05mg/ml albumine. One unit MMP catalyzes the hydrolysis of 1µmol Dnp-peptide/min. at 37°C and pH 7.0. Alternatively the fluorogenic substrate (7-Methoxycoumarin-4-yl) acetyl-Pro-Leu-Gly-Leu-N-β-Dnp-L-(α,β-diaminopropionyl)Ala-Arg-NH2 (Knight et al. 1992) can be used. Hydrolysis of the Gly-Leu bond separates the highly fluorescent (7-Methoxycoumarin-4-yl)acetyl group from the 2,4-dinitrophenyl resulting in an increase of fluorogenic intensity. The Km value for the gelatinase A is 7.0x105M-1s-1. Substrate should be kept as a 9.15mM stock solution in DMSO (10mg/ml). In the assay the substrate concentration should be ~25µM. The assay can be performed in a 96-well microtiter plate (100/200µl per well) suitable for fluorogenic measurements (Ex 328 nm; Em 393 nm). Activation: Requires activation by 2mM (final concentration) APMA or 1mM mersalyl acid for 60-120 min. at 37°C. We do not recommend to use trypsin for activation! Do not dilute enzyme for activation! Inhibitors: Activated enzyme is inhibited by tissue inhibitors of matrix metalloproteinase-2 (TIMP-2) and by chelators of divalent cations like EDTA or o-phenanthroline. |
| Regulatory Status: | RUO - Research Use Only |
