Bio-Rad Ready Gel Precast Gels for 2-D Electrophoresis - Manual (Page 22)

Section 7
Protease Analysis by Zymogram PAGE
7.1 Introduction
Ready Gel zymogram gels are used to test for proteolytic activity when performing protein characterizations.
Gels are cast with gelatin or casein, which act as substrates for proteases that are separated on the gel.
Proteases are detected by renaturing the enzyme followed by a development period in which the protease
breaks down the substrate. Zymogram gels are stained with Coomassie Blue R-250, which stains the
substrate while leaving clear areas around active proteases.
7.2 Ready Gel Zymogram Gel Composition
Gel buffer 0.375 M Tris-HCl, pH 8.6
Cross-linker 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 12 weeks from the date of manufacture
7.3 Ready Gel Zymogram Gel Selection Guide
Ready Gel zymogram gels are available with either gelatin or casein as substrate and should be selected
based on their substrate and separation range.
Zymogram Gel Optimal Separation
10% zymogram gel with gelatin 30–150 kD
12% zymogram gel with casein 20–120 kD
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