ThermoFisher Scientific E-PAGE Gels, Technical Manual

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46
Silver Staining Methods,
Continued
SilverQuest ™
Silver Staining
Protocol
A brief protocol for staining one E-PAGE ™ Gel with the SilverQuest ™ 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 microwave safe container such that the
gel moves freely during all staining and washing steps.
Results obtained with SilverQuest
™ Silver Staining Kit are shown on page 58.
Step Reagent Fast Procedure
Fix Ethanol 80 mL
Acetic Acid 20 mL
Ultrapure water 100 mL
Final Volume 200 mL
Add 150 mL, microwave for
45 seconds.
Agitate the gel for 1.5 hours.
Wash Ethanol 60 mL
Ultrapure water 140 mL
Final Volume 200 mL
Add 150 mL, microwave for
45 seconds.
Agitate the gel for 10 minutes.
Sensitize Ethanol 30 mL
Sensitizer 10 mL
Ultrapure water 60 mL
Final Volume 100 mL
Add 100 mL, microwave for
30 seconds.
Agitate the gel for 15 minutes.
First
Wash
Ultrapure water 150 mL Add 150 mL, microwave for
45 seconds.
Agitate the gel for 10 minutes.
Second
Wash Ultrapure water 150 mL Add 150 mL, microwave for
45 seconds.
Agitate the gel for 10 minutes.
Stain Stainer 1 mL
Ultrapure water 99 mL
Final Volume 100 mL
Add 100 mL, microwave for
30 seconds.
Agitate the gel for 1.5 hours.
Wash Ultrapure water 100 mL Add 100 mL, 1 minute.
Develop Developer 10 mL
Developer Enhancer 1 drop
Ultrapure water
90 mL
Final Volume 100 mL
Add 100 mL, 8–10 minutes.
Stop Stopper 10 mL Add 10 mL directly to Developing
solution, 10 minutes.
Wash Ultrapure water 100 mL Add 100 mL, 10 minutes.