protein elution - any idea to elute protein (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.

regards
alie

Electroelution should do the work, but you need to have equipment to do that.
http://www.protocol-online.org/cgi-bin/prot/jump.cgi?ID=1514

If you need protein for sequencing you can use Westernblotting.

For preparative purification by molecular weight I would rather try size exclusion chromatography (gel filtration).

hi

electroelution can do the work as mentioned before, i am working with it.

but if u want to use very cheap method, u can just cut the protein band from the gel (after using commasie). put

the band in a tube and add 1-3ml of protein running buffer and just leave it for couple of days (i left it over the

weekend). u will see that the protein diffuse in a passive way to the buffer and it become bluish. u will find most of ur

protein in the buffer.

have fun

Yes, that could work. I would cut gel band to small pieces (or even grind to paste) and extract at least two times.

Thanks for giving suggestion....i have done size exclusion but i am not able to resolve these proteins they are close to each other.....

so i thought to extract protein from SDS PAGE.

after electro elution can i use for HPLC to check purity?