Zymogram gels

Invitrogen Novex Zymogram Gels are excellent tools for detecting and characterizing proteases that utilize casein or gelatin as a substrate.

Interactive selection toolAvailable Novex Zymogram Gels

Migration chart

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) 
Storage conditions 2–8°C
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
Post-staining required Yes Yes No
Separation range 20–120 kDa 30–150 kDa 10–220 kDa
 
Migration chart

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)

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) 
Storage conditions 2–8°C
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
Post-staining required Yes Yes No
Separation range 20–120 kDa 30–150 kDa 10–220 kDa
 
Migration chart

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)