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Our Abpromise guarantee covers the use of ab157344 in the following tested applications.
The application notes include recommended starting dilutions; optimal dilutions/concentrations should be determined by the end user.
≥1’400mU/mg protein (Y. Masui, et al.; Biochem. Med. 17, 215 (1977)). One unit is defined as the amount of enzyme that hydrolyzes 1µmol 2,4-dinitrophenyl-Pro-Gln-Gly-Ile-Ala-Gly-Gln-D-Arg-OH per min. at 37°C, pH 7.0.
Precursor enzyme needs activation using 2 mM APMA (aminophenylmercuric acetate) for 60-120 min. or 100 µg/ml TPCK-trypsin for 30 min. at 37°C. If required trypsin can be inhibited by incubation with 1µl of 1 mg/ml aprotinin solution for 10 min. at 25°C. If working with MMP9/TIMP-1 complexes, it is recommended to use stromelysin-1 (MMP3) for activation. Incubation at 37°C for 2 hours at 40:1 (MMP9:stromelysin-1) will remove only the propeptide to give the active form of the enzyme.The active enzyme is inhibited by TIMP1 (tissue inhibitor of matrix metalloproteinase-1) and by chelators of divalent cations like EDTA or o-phenantroline.
Shipped at 4°C. Upon delivery aliquot and store at -80ºC. Avoid freeze / thaw cycles.
Preservative: 0.05% Sodium azide
Constituents: 0.05% Brij, 0.06% Calcium chloride, 0.00001% Zinc chloride, 0.79% Tris HCl, 1.17% Sodium chloride
This product is an active protein and may elicit a biological response in vivo, handle with caution.
This blot was produced using a 4-12% Bis-Tris gel under the MOPS buffer system. The gel was run at 200V for 60 minutes before being transferred onto a nitrocellulose membrane at 30V for 70 minutes. The membrane was blocked for an hour before being incubated with anti-MMP9 antibody [EP1254] (ab76003; 5 microgram per mL) at 4°C overnight. Antibody binding was detected using infrared labelled goat anti-rabbit (green) antibody (diluted 1:20000) for 1 hour at room temperature before imaging.
Please note: All products are "FOR RESEARCH USE ONLY AND ARE NOT INTENDED FOR DIAGNOSTIC OR THERAPEUTIC USE"