I have been running blue native gels for my chloroplast samples, the problem is that the gel won't run completely. I have seen gel pictures in different papers that the whole sample runs really nicely but in my case the bottom part of the sample never runs and I have not manage to see the bottom bands of my samples. This is how my gel looks.
No matter how much I increase the voltage the gel doesn't run more.
notes:
1.I use 2 different cathode buffers, 1/3 of the gel with the cathode buffer containing coomassie and the rest without coomassie.
2.My anode buffer has no Tricine
3.I use Coomassie 250G
this pattern looks like too much salt in the samples. Can the sample be dialysed or the salt removed before loading?
Thanks for the replies, I will check the pH of the samples, as I'm sure with the pH of the buffers but I will recheck them also.
I run the gel in cold room (4 degrees) overnight, for the first 5-6 hours I run it at 9 mA and just before leaving the lab I turn it to 5 mA to run the whole night so this gel is the gel after almost 24 hours of running!!!!
I make the gradient gel myself, the gel above is 6-14% gel.
I will also try to remove the salt from the samples. I hope all these would help.
Thanks again
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