horrific western blots , please help - (Jul/11/2010 )
hi everyone, i have been working on these western blots and am seeing these horrible blots. We run westerns routinely in the lab using pre-cast gels (150V for 140 min) and there have been no problems thus far. also, please note that i previously ran the same sample under identical conditions and everything was hunky-dory. but now all i see are these squiggly bands that start out okay but go all loopy/distorted as the proteins progress through the gel. finally the standards run just fine!!
i'm at my wits end with these. any advice will be greatly appreciated
many thanks.
It looks like there is too much salt in the loaded sample, which can result in the bands getting distorted like that, though this doesn't account for your good result with the same samples, unless you used a different batch of loading buffer or something. It could also be that the pre-cast gels are going off or the the buffer composition not being quite right (especially if you re-use buffer).
bob1 on Jul 11 2010, 03:26 PM said:
hi thank you for your prompt reply. unfortunately, i used loading buffer from the same batch. gels dont expire until september and I ran the samples that worked well a day before these, hence buffer composition change is an unlikely suspect..
airbubbels at the bottom of your plate?