Hi all,
I read many protocols and Q&A in ResearchGate. Many can run protein SDS-page electrophoresis at 200V constant using same set of equipments that I use (Bio-Rad Mini-PROTEAN® Tetra Cell Systems). What are the conditions that they use to run 200V?
I tried once 150V for about 1 hr by using fresh cold running buffer (stored at 4°C). When the protein sample was running for about half a hour, the chamber became very hot, even in the cold room, and the gel started melting.
Is there a method to determine the maximum voltage for gel electrophoresis as well as for western blot of proteins? Can I determine it by looking at current and resistance values on the power supply unit?
Thank you.
Did you check the current generated through your gel while applying 200 V ? With our Bio-Rad set-up (Miniprotean III), which is abot 8 cm across and 0.75 mm thick, running Lämmli gels (12 % acrylamide) yields abot 40 mA at the beginning and 18-20 mA at the end of the run. Of course, these values will depend on the thickness of the gel. It is also possible that your buffer concentration is wrong; check the original recipe.
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