Hello,
For SDS-PAGE western blots: do you usually run your gels on ice or room temp.? For example if I use these conditions: Stacking 70V 30min, then separating 120V for 2 hrs. Is it okay to run it on ice? Or it is bad because of SDS inside my running buffer?
Thank you!
Hi Olga, We run the gel at room temperature . For staking 50v and for resolving 100 V. No need to run on ice for Western blot, it is required for enzymes assay.
We also run SDS-PAGE at RT at even higher voltages (150 V stacking and 200 V separating). Never had any problems.
Thanks! I always run on ice. Recently somebody told me that running on ice is bad. Is that true?
hi olga,
i do not use ice per se but i do keep my tank cool at around 15-20 C. That has done a lot of good for my gels. Also the running conditions should be 50V through stack and 90-100 V in resolving. It will take some time but its worth.
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