Activity Analysis by Zymography
Adapted for Matrix Metalloproteases (MMPs)
Technique
Zymography is a useful qualitative tool for detection and analysis
of the level and type of the proteases expressed in different cell types/tissues at any given time and/or after different treatments. This technique, known mostly as gelatine / casein zymography, identifies enzymes activity in biological samples using sodium dodecyl sulfate (SDS)-polyacrylamide gels impregnated (copolymerized) with gelatine / casein. To maintain enzymatic activity, the samples are electrophoresed under nonreducing conditions. After removal of the SDS from the gel by Triton X-100 and incubation in a calciumcontaining buffer, the partially renatured enzymes can degrade the copolymerized substrate, leaving a cleared zone that can be detected after staining of the gel. In the presence of SDS, the enzymes are denatured exposing their active site, which permits both the latent and active forms of MMPs to exhibit its activity after the partial renaturation.
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Advantages
• Simple tool for separation and identification of second level structural gene products
• Detection of enzymes and their structural isoforms
• Ideal for screening drugs and examination of protease inhibitors
• Very low sample amount require
• Cheap method and easy to handle
Our Service
• Send us your samples together with a completed order form
• In 1.5 working days after sample arrival we provide an image of zymography per E-Mail
• Performance of zymography with marker and positive control (MMP2)
• Storage of sample(s) for 2 months
Please find here the order form with guidelines or contact us directly.