protein elution from SDS PAGE - any idea how to elute protein from SDS PAGE (Apr/15/2008 )
hello protein friends,
i am new to this protein world i have a problem........i have to elute my protein from SDS PAGE .....is any body having any idea how to elute protein from SDS PAGE.i need a conventional method i cant afford for any kits now....
i hope you people will help.
What do you want to do with the protein afterwards? I can think if a few reasons to purify a protein by electrophoresis, and a few ways to do it.
The simplest would be to cut out the band and place it in dialysis tubing, and re-electrophorese. Visualizing the band before excising will require picking the right dye and avoiding conditions that would fix the protein (methanol, acetic acid). Not sure if ponceau would work. Also, choose a low MW cut off for the tubing because SDS will denature the protein, making it easier to slither through the pores.
If you want to get aa sequence info from the protein, check with the group performing this activity. When I have done this, the mass spec facility wanted the protein transferred to a PVDF membrane (sending them the membrane slice).
If the protein is to be used as an immunogen, you may be able to give the gel slice to the animal facility. But it is no problem to raise antibodies to an impure protein and select for specific mAbs or affinity purify antisera later.
I guess it matters what the protein will be used for. Using SDS PAGE as a purification step is not ideal in most cases.