1 introduction, 2 criterion zymogram gels, 1 gel composition – Bio-Rad Criterion™ TBE-Urea Precast Gels User Manual
Page 29: 2 gel selection guide, 3 zymogram buffers, Protease analysis (chapter 8), Protease analysis by zymogram page

Protease Analysis by
Zymogram PAGE
8
8.1 Introduction
Criterion
™
zymogram gels are used to test for proteolytic activity. Gels are cast with gelatin or casein,
which acts as a substrate for proteases that are separated in the gel under nonreducing conditions.
Proteases are detected by first renaturing the enzymes and then allowing them to break down the
substrate. Zymogram gels are stained with Coomassie Brilliant Blue R-250 stain, which stains the
substrate while leaving clear areas around active proteases.
8.2 Criterion Zymogram Gels
8.2.1 Gel Composition
Gel buffer
0.375 M Tris-HCl, pH 8.6
Crosslinker
2.6% C
Stacking gel
4% T, 2.6% C
Storage buffer
0.375 M Tris-HCl, pH 8.6, 0.2% NaN
3
Shelf life
~8 weeks at 2–8°C; expiration date is printed on each cassette
8.2.2 Gel Selection Guide
Zymogram Gel
Optimum Separation Range
10% with gelatin
30–150 kD
12.5% with casein
20–120 kD
8.3 Zymogram Buffers
See Appendix B for buffer formulations. Do not adjust pH unless instructed to do so.
Running buffer (1x)
25 mM Tris, 192 mM glycine, 0.1% SDS
Dilute 100 ml 10x stock (catalog #161-0732) with 900 ml diH
2
O
Sample buffer
62.5 mM Tris-HCl, pH 6.8, 4% SDS, 25% glycerol, 0.01% Coomassie
(catalog #161-0764)
Brilliant Blue G-250
Technical Support: 1-800-4BIORAD • 1-800-424-6723 • www.bio-rad.com
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- Criterion™ TBE Precast Gels Criterion™ Cell Criterion Precast Gels Criterion Dodeca Cell 2-D Electrophoresis Workflow Criterion™ IEF Precast Gels Criterion™ Zymogram Precast Gels Criterion™ XT Tris-Acetate Precast Gels Criterion™ Tris-Tricine Precast Gels Criterion™ XT Bis-Tris Precast Gels Criterion Stain Free™ Tris-HCl Gels Criterion™ Tris-HCl Precast Gels Criterion™ TGX Stain-Free™ Precast Gels Criterion™ TGX™ Precast Gels