Hy,
if you want to label intracellular epitopes you need to perform this procedure, if you want to label cell surface proteins/markers you don't.
Usually you can pretreat tissue slices with methanol for 1-10 min at -20°C or you use Triton X100 in the blocking (about 0.05%) and antibody solutions (0.025%).
Good luck
Hi Rahul
To be sure that the antibodies get inside the cells, you should use suitable permeabilised solution like Triton 100X FOR T 10 min to permeablised cell membrane.
The need for permabilization is dependent on the fixative you use. If you fix in ice-cold acetone then you don't need to permeabilize but if you fix in paraformaldehyde then you do. I normally fix cells in 4% paraformaldehyde and then permeabilize for 2 mins in 0.2% TritonX100
Besides using the above stated reagents to permeabilize cells' membrane, you could use an another well known reagent ,saponin.
There are, of course, several different methods which facilitate permeability of cellular plasma membrane. However, such a pre-treatment is unnecessary if you intend to us formalin-fixed and paraffin-embedded tissue or cell specimens.
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