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Immunology on fixed bacteria - What are the basics of fixing cells for immunological analysis? (May/01/2007 )


If I have an exponentially growing culture of a "garden variety" bacterium, say Pseudomonas or some other gamma proteobacterium, how do I fix the cells so that I can ship them to someone who has antibody, and who can then stain them?

The antibody is to a protein that should be expressed and localized in the inner cell membrane. The antibody should recognize the bit of the protein that sticks out of the membrane, in the periplasmic space.

I haven't done this sort of thing, and thought y'all might be able to get me started.


p.s. Does Maniatis cover this sort of thing?


i dont know too much about bacteria but my guess would be use a crosslinker like paraformaldehyde or treat them like a smear (fix and protect using one of those waxy sprays designed for sputum/cervical smears etc).
word of warning tho any fixative you use at your end will change the epitope retrieval at the other and i doubt your bacteria would survive a microwave or pressure cooker no matter what type of glue you used.
you might be better off shipping the antibody and doing the immuno yourself (or there might be a standard protocol for this kind of thing that the next person will tell you blush.gif )