Bio-Rad Image Lab™ Software - Manual (Page 225)

Electrophoresis with Stain-Free Gels
User Guide | 225
Electrophoresis with Stain-Free Gels
Stain-free gels are made and packaged without sodium dodecyl sulfate (SDS),
allowing them to be used for both SDS and native polyacrylamide gel
electrophoresis (PAGE) applications.
To perform electrophoresis with stain-free gels
1. Prepare the sample and running buffers.
2. Set up the electrophoresis cell.
3. Perform the run.
Imaging Gels
Use unstained standards with stain-free gels, as some prestained standards are not
compatible with stain-free technology. To monitor electrophoresis, use a 1:1 mixture
of unstained and prestained standards.
Setting up a protocol for stain-free gels is very similar to setting up protocols for
other applications. Follow the instructions in Setting Up a New Protocol on page 72 .
Choose one of the following activation times based on your sample and the purpose
of your experiment:
 Gels used in blotting — use 1 min activation for optimal results when
performing western blotting followed by immunodetection.
 Good sensitivity — use 2.5 min activation when samples are abundant
and when a fully optimized signal-to-noise ratio is not necessary.
 Best sensitivity — use 5.0 min activation for detection of proteins that are
in low concentration and for the best quantification of the maximum
number of bands. Because the reaction is near completion after 5 min, this
method offers an optimal signal-to-noise ratio.
Note: If the gel has already been activated for 2.5 min, activating it for another
2.5 min might improve it. But activating an image for more than 5 min will not.