Tween-20 vs Triton-X vs Saponin - (Nov/29/2011 )

Does anyone know how different or similar all three detergents are in terms of using it in our flow buffers?

I know Triton is stronger/harsher vs Tween.

And would you have a separate permeabilisation step with detergent and then remove detergent from the antibody incubation buffer & wash buffer?

Apparently it keeps the membrane pores open? or anyone know the mechanism of action of detergents in immuno?

We use saponin in our lab. For intracellular staining, saponin makes pores in the cell membrane, allowing your antibody to enter the cells and stain the intracellular target. So you first need a permeabilization step, and then you add the antibody in the presence of this permeabalizing buffer, as the membrane pores will close up in the absence of saponin preventing your antibody from entering otherwise. Once you have done the intracellular staining though I dont see a reason why you need to have the detergent any longer.

Unfortunately I have no experience using another detergent though.