no front in sds-electrophoresis - no front is formed on my gel - why? (Oct/05/2009 )
I've recently prepared the new solutions for my WB, and now, as I've loaded my probes on the gel they did not form the front with the bromophenol blue. Never seen anything alike before. The prestained markers are running the way they should, though it seems to be a lilttle bit too quick. I've checked the pH of all the buffers - it's ok.. but the BB in the probes goes much higher than the electrophoresis front (which is stainless and almost invisible) as separate bands... so, I'm wondering - shoul I do the transfer or abandom the whole thing? And what could be wrong in there?
Thanks for your hints in advance!!
as long as the standards look okay, it would be worth trying the transfer (although, if you have enough sample, you may want to stain the gel instead).
as for the cause(s). i would check the buffers again. the pH may be okay (although checking the pH with sds present may be a problem) but the concentration of the components may be off or something extra may have been added or something may have been omitted.