b'Zymogram gelsEasy in-gel protease analysisInvitrogen Novex 10% Zymogram Plus (gelatin) gels areSpecificationsuseful for the detection and characterization of proteases Shelf life: 2 monthsthat use gelatin as a substrate. The proteases are run under denaturing conditions and visualized as clear bands Average run time: 90 minutesagainst a dark background using a simple renaturing, Separation range: 20220 kDa (Figure 22) developing, and staining protocol. Zymogram gels are commonly used to detect matrix metalloproteases. Novex Polyacrylamide concentrations: fixed 10% (with gelatin) 10% Zymogram Plus gels are highly sensitive, detecting as little as 5 x 10 6units of collagenase.Gel dimensions: 8 x 8 cm (1 mm thick) Maximum sample volume per well: 20 LNovex Zymogram Plus (gelatin) gel characteristics.Zymogram Plus gelatin gelGel composition 10% Tris-glycine gelSubstrate 0.1% gelatinSensitivity 5 x 10 6units of collagenasePost-staining required? YesSeparation range 20220 kDaGood to knowHow do Novex zymogram gels work?Protease samples are denatured in SDS buffer under nonreducing conditions and without heating, and run on a Novex Zymogram Plus gel in Tris-Glycine SDS Running Buffer. After electrophoresis, the proteases are renatured by incubating the gel in Invitrogen Novex Zymogram Renaturing Buffer, which contains a nonionic detergent. The gels are then equilibrated in Invitrogen Novex Zymogram Developing Buffer to add divalent metal cations required for enzymatic activity, and then stained and destained. Regions of protease activity appear as clear bands against a dark blue background where the protease has digested the substrate.Learn more at thermofisher.com/zymogram30'