ThermoFisher Scientific E-PAGE Gels, Technical Manual

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47
Silver Staining Methods,
Continued
SilverXpress ®
Silver Staining
Protocol
A brief protocol for staining one E-PAGE ™ Gel with the SilverXpress ® Silver
Staining Kit is described below. For details, refer to the product manual available
at www.lifetechnologies.com or contact Technical Support (page 72).
After electrophoresis, remove the gel from the cassette (page 24). For all staining
and washing steps described below, be sure to use sufficient volume of reagent
to completely cover the gel in a suitable container such that the gel moves freely
during all staining and washing steps.
Results obtained with SilverXpress
® Silver Staining Kit are shown on page 59.
Note: The final volumes of solutions containing both methanol and water reflect
a volume shrinkage that occurs when these two reagents are mixed. Do not
adjust volumes of components or final volume.
Step Reagent Procedure
Fix Ultrapure water 270 mL
Methanol
300 mL
Acetic Acid 60 mL
Final Volume 600 mL
(see note above)
Fix the gel in 3 changes (~200 mL
each) of fixing solution for 90
minutes. Change the fixing
solution every 30 minutes.
Sensitize Ultrapure water 105 mL
Methanol
100 mL
Sensitizer 5 mL
Final Volume 200 mL
(see note above)
Incubate the gel in 2 changes of
Sensitizing Solution for 1 h, each.
Decant the solution.
Wash Ultrapure water 200 mL Wash the gel twice with ultrapure
water for 15 minutes each.
Stain Stainer A 5 mL
Stainer B 5 mL
Ultrapure water 90 mL
Final Volume 100 mL
Incubate the gel in Staining
Solution for 30 minutes. Decant
the solution.
Wash Ultrapure water 200 mL Wash the gel twice with ultrapure
water for 10 minutes each.
Develop Developer 5 mL
Ultrapure water 95 mL
Final Volume 100 mL
Incubate the gel in Developing
Solution for 1–5 minutes.
Stop Stopping Solution 5 mL
Add the Stopping Solution
directly to the Developing
Solution when the desired
staining intensity is reached.
Incubate the gel in Stopping
solution for 20 minutes. Decant
the solution.
Wash Ultrapure water 200 mL
Wash the gel three times with
ultrapure water for 20 minutes
each.