Protein Denaturation

[noezn]

My doctor father wants me to show, that the upper band of the two bands I get in my Blot is a dimer obf the lower one.
This complex is very stable,as it is not diminished by SDS-PAGE and ß-mercapto. SO I tried to do an urea denaturation but then I got no signal in my Blot anymore.
Has anyone an idea of how I can show this ? I would be very happy for every answer

[admin]

Hello Noezn,
that is an interesting question.

Have you tried different concentrations of beta-mercaptoethanol?


Also, if it is a dimer of two proteins it should be an ionic interaction
which should be broken by boiling + salt/sds/beta-mercapthoethanol.

If not, then you should look towards using more harsh methods or analysing the bands after specific digestion.