Bubbles in western blot gel "AFTER" running gel - (Sep/16/2015 )
So I typically run western blots with no problem. However, during my most recent experiment something happened which I can't figure out the reason for.
So I loaded my samples into my gel like always. The gel seemed fine and everything went as usual. I ran the gel at 120V for 2 hours. However, upon removing the gels I saw that both of the gels were full of bubbles which didn't exist before running the gel.
I'd appreciate it if someone could let me know why this happened and how to prevent this from happening again.
Thanks a lot for your help.
Were you using precast gels or did you make them? First guess would be that the gels were not fully solidified if you made them and so the bubbles formed when the current is being passed through the gel. Also make sure you remember to equilibrate the gels in running buffer to room temp before running the gel.
this was probably due to heat generated during the run causing dissolved gases to be released.
if you pour your own gels then you may want to degas the solutions prior to addition of the catalyst and initiator. at the very least, make sure all of your solutions are at room temperature before formulating your gel.
if the gels are purchased precast then you should ensure that they are warmed to room temperature before opening the package. the cold gel can absorb gases which will be released when warmed.
or, the gels overheated during the run. this can happen if the electrode buffers are improperly formulated or if the power supply is set incorrectly or the safeguards fail.