I am new to western blot and when I was running it today, the machine stopped the run by itself and I realized it only one hour after. The ladder was barely separated, so I restarted the run until I saw the appropriate ladder separation for my experiment, which was roughly an additional hour. I am very concerned that the interruption disrupted the western blot and protein migration. Is this a really big issue? Thank you.
That should not be a big problem, since you caught it relatively quickly. Without any electrical current to push the migration, I would think they'd be pretty stable in the gel. Since the ladder separation looked appropriate, I would think the rest of the wells ran correctly too! Keep us updated on the results. A big disadvantage with many drawn-out lab techniques like Western Blotting is that you don't know what impact any small alterations might have on the final product until the very end (usually the next day for us)... it makes for a frustrating 18-24 hrs! haha. Good luck!
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