Zymogram Gel Staining
Prior to staining zymogram gels, a sample proteases must
be first be allowed to renature and the allowed to 'break
down' the substrate contained in the gel.
Zymogram Renaturing Solution:
2.5% TritonX-100
Zymogram Development Solution:
50 mM Tris
200 mM NaCl
5 mM CaCl2 (anhydrous)
0.02% Brij-35
Adjust to pH 7.0
Zymogram Staining Solution:
40% methanol
10% acetic acid
0.5% Coomassie Blue R-250
Zymogram Destaining Solution:
40% methanol
10% acetic acid
Place gels in renaturing solution fo 30 min at RT. Incubate
gels in development solution at 37°C for a minimum of 4 h.
Highest sensitivity is typically achieved with overnight
incubation. Optimal results should be determined
empirically. Stain gels with Coomassie Brilliant Blue R-250
staining solution for at least 1 h @ RT. Destain until clear
bands appear against the blue background, approximately
30-60 min.
Prior to staining zymogram gels, a sample proteases must
be first be allowed to renature and the allowed to 'break
down' the substrate contained in the gel.
Zymogram Renaturing Solution:
2.5% TritonX-100
Zymogram Development Solution:
50 mM Tris
200 mM NaCl
5 mM CaCl2 (anhydrous)
0.02% Brij-35
Adjust to pH 7.0
Zymogram Staining Solution:
40% methanol
10% acetic acid
0.5% Coomassie Blue R-250
Zymogram Destaining Solution:
40% methanol
10% acetic acid
Place gels in renaturing solution fo 30 min at RT. Incubate
gels in development solution at 37°C for a minimum of 4 h.
Highest sensitivity is typically achieved with overnight
incubation. Optimal results should be determined
empirically. Stain gels with Coomassie Brilliant Blue R-250
staining solution for at least 1 h @ RT. Destain until clear
bands appear against the blue background, approximately
30-60 min.
