I have tried using BN-PAGE for mitochondrial complexes isolation. I have followed several protocols (Nature protocols-2006, JOVE-2017). But I experienced a problem with running the gel. The dye front is not moving out of the wells. Also, I am not using gradient gels. Could that be an issue? And are there any precast gels available for BN-PAGE (using imidazole and not bis-tris) commercially?
Dear Gowthami,
I believe that non-gradient gels could be fine for BN-native unless you are concerning about different complexes (eg super complex). However, we have been using NativePAGE Bis-Tris 3-12% precast gels from thermo which results a good separation. Just double check gel and buffer compositions as well as mito prep. Good luck
Hi,
Thank you for the thought. I will try the gradient ones. One more question. The protocol that I am following is using imidazole and tricine instead of Bis-tris (Jove protocol 2017). So will it be suitable to use the NativePAGE Bis-Tris gels or else should I change the protocol which goes with Bis-Tris in the buffer system? Thanks in advance!
Hi!
People are using imidazole tricine and Bis-tris in buffers and gel, respectively or vis versa. Better to try and let me know the outcome. Good luck.
Hi Gowthami!
Could be useful to you. https://www.sciencedirect.com/science/article/pii/S2666166720300769
Cheers
- Badges
- Science topic
- Similar topics
- Gels