Bio-Rad Ready Gel Precast Gels for 2-D Electrophoresis - Manual (Page 30)

10.2 Peptide Gel Staining
Peptides and small proteins are prone to diffusion and loss during staining. The following protocol uses a
fixation step to prevent sample loss and is suitable for detection of bands as low as 10–20 ng.
Fixative Solution
40% methanol
10% acetic acid
Coomassie Brilliant Blue G-250 Stain Solution
0.025% Coomassie Blue G-250
10% acetic acid
Destain Solution
10% acetic acid
Place gels in fixative solution and equilibrate for 30 min. Stain gels with Coomassie Brilliant Blue G-250 stain
solution for 1 hr. Stain should only be used once. Reuse of stain could result in loss of sensitivity. Destain
gels 3 times for 15 min or until the desired background is achieved. Some peptides may not be completely
fixed and may diffuse out of the gels if fixing and staining times are greatly exceeded.
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