SDS-PAGE Dimer! - (May/23/2006 )

Hi,everyone:

i was doing some Western Blotting(SDS-PAGE) to test poly-antibodies against some transcript factors.always there were some high-weight bands,about double MW of the monomer.somebody else tell me they are dimers.

i need a perfect result to show only one correct band at right MW,

So,Is there any methods to remove the dimer?
should I change the conditions of treating samples? for example , boiling to 60 degree C and cool down slowly?
Or doing the non-reduced PAGE?
Or subcelluar separation ?
Or change the detergent,for example ,Triton X-100?


And ,i don't think it was glycoprotein,although many dimers are found glycosylated,it is unusual to found transcript factors glycosylated,isn't it?


Thanks for any response

May be you did'nt reduce well your protein, that's why you still have dimer.
You might prepare fresh loadiing buffer,and boil 5 minutes (90-95°C).
Usually people prepare their buffer with 1% beta-mercaptoethanol, but some proteins have quite a lot of cystein, and you should then increase to 5%.