permeabilization without killing cells

Howard Shapiro hms at shapirolab.com
Tue Oct 30 11:17:50 EDT 2007


Anuraag Sarangi wrote:

>  
> we have an antibody that detects an intracellular epitope of the target
> protein.  We have good separation of signal over isotype-matched
> background when we use intracellular staining by permeabilizing and
> fixing the cells.  What we could like to know is whether there is any
> way that we can use the antibody for FACS sorting without killing the
> cells by permeabilization?  We would like to collect the sorted, viable
> cells and culture them.
>  

See: Berglund DL, Starkey JR: Introduction of antibody into viable cells
using electroporation. Cytometry 1991; 12:64-67. This is free online.

This subject probably deserves another look given the current level of
interest in demonstration of cell signaling proteins, cytokines, etc. It
would be nice to be able to sort live cells and follow them after
staining intracellular material, keeping in mind that the analytical
procedure may perturb the system.

-Howard



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