Novex Zymogram Gels are excellent tools for detecting and characterizing proteases that utilize casein or gelatin as a substrate. Casein and gelatin are the most commonly used substrates for demonstrating the activity of proteases. Novex Zymogram Gels can be used to analyze a variety of enzymes, including matrix metalloproteinases, lipases, and other proteases. Novex Zymogram Gels are based on tris-glycine gel chemistry containing gelatin or casein as the substrate.
How Novex Zymogram Gels work
Protease samples are denatured in SDS buffer under non-reducing conditions and without heating, and run on a Novex Zymogram Gel using tris-glycine SDS running buffer. After electrophoresis, the enzyme is 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.
|Available gel sizes||Mini: 8 cm x 8 cm (1 mm thick)|
|Shelf life||2 months|
|Gel chemistry||Tris-glycine–containing gelatin or casein|
|Running buffer||Tris-glycine SDS running buffer|
|Available polyacrylamide concentrations||10% (w/gelatin), 12% (w/casein), 4–16% (with blue casein)|
|Separation range||10–220 kDa|
|For use with (equipment) mini gels||Mini Gel Tank or XCell SureLock Mini-Cell|
|Mode of separation||Molecular weight|
|Zymogram Gelatin Gel||Zymogram Casein Gel||Zymogram Blue Casein Gel|
|Gel composition||10% tris-glycine gel||12% tris-glycine gel||4–16% tris-glycine gel|
|Substrate||0.1% gelatin||0.05% casein||0.1% casein, with blue stain incorporated in gel|
|Sensitivity||5 x 10-6 units of collagenase||7 x 10-4 units of trypsin||1.5 x 10-3 units of trypsin|
|Separation range||20–120 kDa||30–150 kDa||10–220 kDa|
The numbered bands refer to the following proteases:
Band 1: Collagenase Type I (140 kDa)
Band 2: Thermolysin (37 kDa)
Band 3: Chymotrypsin (30 kDa)
Band 4: Trypsin (19 kDa)
For Research Use Only. Not for use in diagnostic procedures.