Flow cytometry - permeabilization protocol? - (Sep/06/2005 )
Hi! I am in need a protocol for permeabilizing cells for flow cytometry that uses a homemade recipe rather than a commercial permeabilization buffer. Any help is most appreciated!
Fix your cells in 2% paraformaldehyde then permeabilise in 0.1% Triton-X100/PBS. My colleague used this in his recent paper and it works well. The paper is in the in press sections of FEBS letters if you want to have a look at the results.
Mutation of the extracellular domain of tumour necrosis factor receptor 1 causes reduced NF-kappaB activation due to decreased surface expression Accepted Manuscript, 31 August 2005
Siebert S, Fielding CA, Williams BD, Brennan P
Full Text on ScienceDirect
Alternatively you could use saponin (not sure what percentage to use), but it needs to be in all staining and wash buffers as the effect is reversible.
Hope this is off some use,