permeabilization - (Jun/27/2006 )

A problem about Immunostaining:

After fixation of the cell,I want to permeabilize them beacause my target protein may expressed intracellularly. If I utilize Triton X-100, what is the optimal concentration? The cell in my experiment are HEKs(human embryonic kidney(cells).

Additionally,I wondering if the Triton appropriate for the use of membrane-associated antigens.

Thx!

I use 0,2% Triton-X100 in PBS++ for 25'/RT for most of my cell lines. It usually works but it could be you would have to adjust the time

I use 0.2% triton 2-5 min RT or acetone (10 min in -20C). Anyway u have to optimise the time.