Red Blood Cell (Hypotonic) Lysis Buffer

Warren Gallin wgallin at gpu.srv.ualberta.ca
Tue Jun 23 19:40:39 EST 1998


In Article <35904CCE.12A5 at ktb.net>, Bert Gold <bgold at ktb.net> wrote:
>Colleagues,
>
>For a DNA extraction at work, it would be good to be able to mix up
>a hypotonic buffer capable of lysing red cells, but keeping white 
>cells (or at least nuclei) intact.  Anybody know a recipie for
>such a salt-water mix?

When making hybridomas it is desirable to lyse the RBCs and leave the
lymphocytes viable.  We resuspend the cell pellet from a single spleen in 5
ml of ice cold 0.75% ammonium chloride, on ice, five minutes, then pellet
the cells, discard the now red supernatant, and resuspend the cell pellet in
normal medium, or buffered saline.  Virtually all the RBCs are gone and the
lymphocytes are still viable.
Warren Gallin
Department of Biological Sciences
University of Alberta
Edmonton,  Alberta     T6G 2E9
Canada
wgallin at gpu.srv.ualberta.ca



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