Developing Gel instead of PVDF - (Mar/16/2006 )
Hello, has anyone every developed a gel instead of using the PVDF? I am thinking I am losing a lot of my protein whenever I transfer to PVDF. If so could you please give me details of how to do it.
"developed" for or with what?
I've probed gels for glycoproteins, ca-binding proteins, etc. but antibodies are large and don't infiltrate gels very well, they may only detect proteins near the surface, thereby making them largely unsuitable for gel staining.
Pierce sell some kind of kit for in gel detection. I don't know anyone who has tried it.
Maybe optimising your transfer would be worth considering though. Why do you think you are losing your protein? Are you pre-wetting your PVDF with methanol and then transfer buffer? Can you transfer for longer if your protein is still in the gel?
All the best,
Ceri
Maybe optimising your transfer would be worth considering though. Why do you think you are losing your protein? Are you pre-wetting your PVDF with methanol and then transfer buffer? Can you transfer for longer if your protein is still in the gel?
All the best,
Ceri
I transfer for 3 1/2 hrs on ice, and I do pre-wet my PVDF with methanol and then transfer buffer. I fell that my result are varying from experiment to experiment. Plus, I had heard about developing on the gel but didn't know if anyone had tried it and had any comments on it.
Thanks