BabcockHall Posted Monday at 06:42 PM Share Posted Monday at 06:42 PM I have cast and used SDS PAGE with Tris/glycine/SDS electrode buffer. However, I was planning to use some precast, gradient gels for the first time. I am trying to contact the manufacturer, but I have not yet heard back. "BisTris" is in the product description, and the catalog page mentions the possibility of ordering MES or MOPS buffer. My first question is whether I should change my recipe for the sample loading buffer. My old recipe includes Tris (pH 6.8, same as stacking gel), SDS, 2-mercaptoethanol, glycerol, and bromphenol blue. My second question is what sort of power settings should I use? In the past I have typically used about 100 volts during the stacking portion of the run, which changes to 150 volts or more volts once the samples enter the resolving gel. Link to comment Share on other sites More sharing options...
CharonY Posted Monday at 09:58 PM Share Posted Monday at 09:58 PM I generally do not use precast gels, so I don't have first-hand experience. But I believe that manufacturer generally recommend (obviously) their own loading buffer (LDS). While it is generally recommended to use the same buffer as the running buffer, I believe that most LDS buffer composition are also simple Tris-HCl at pH 6.8. I.e. it shouldn't make much of a difference either way. For power settings, I generally run at constant current as I prefer controlled and more homogenous separation (mostly as I approach this from a 2D perspective). But if your gel are the same size, you can keep the values the same (i.e. 10-15 V/ cm). But depending on the buffer the run time would change. 1 Link to comment Share on other sites More sharing options...
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