I've lysed some cells for Co-IP. For this purpose the lysis buffer is without denaturing detergents. For western blot I now need to add SDS or another appropiate detergent. Can I just add laemmli buffer to the Co-IP buffer, or can I add SDS alone to the CO-IP buffer?
Thanks in advance!
Lämmlli buffer will provide mercaptoethanol for the reduction of disulfide bridges, both intra and intermolecular, as well as the glycerol needed to ensure that the samples will go down into the loading wells of the gel. Otherwise, it also provides the right pH and ionic strength for proper stacking in the stacking gel.
Laemmli buffer contains SDS (anionic surfactant), thiol agent (reduce the disulphide bonds), glycerol, and a color agent. So, you can add it to provide all these reagents.
If you add SDS alone, you need to add all the other reagents.
Just for context: I first thought that maybe, because of the lysis buffer, it wouldnt be necessary to add the "whole" laemmli buffer, since it already has some detergents. But since the lysis buffer leaves the proteins more or less untouched, you should still add the entire laemmli.
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