Permeabilization of cells for FACS staining

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milimandal
milimandal's picture
Permeabilization of cells for FACS staining

Hi,

For intra-cellular staining I permeabilize cells with permeabilizing buffer provided by ebiosciences. I was wondering what other people use for permeabilizing the cells? Can any detergent be used? Permeabilization buffer from ebiosciences is little expensive and I am not sure if we need to spend money on that if it can be done by some cheaper detergent.

Please let me know.

hawknatalyn
hawknatalyn's picture
I am also trying to make my

I am also trying to make my own buffer in lieu of buffer provided by a manufacturer. I came across a recipe for fixing cells by resuspending in final concentration 2% formaldehyde for 10 min. Then permeabilization with 90% methanol on ice.

I have since found a more detailed protocol in J Immunol Methods 315:37-48 which uses: 2% paraformaldehyde fixation for 5 min at RT; then permeabilization: 150 mM NaCl, 5 mM EDTA, 50 mM Tris-HCl, pH 7.4, 0.05% NP40, 0.25% lambda-carrageenan, 0.02% NaN3. This was for simultaneous characterization of intracellular proteins and DNA content

Jen_Floyd
Jen_Floyd's picture
I used to permeabilize cells

I used to permeabilize cells with .1% tween. They were adhered to a slide but I imagine it would work for cells in buffer as well.