Lysis of erythrocytes / streptavidin conjugation with "low backgrounds"

From: Joern E. Schmitz (jschmitz@caregroup.harvard.edu)
Date: Fri Jul 09 1999 - 16:31:15 EST


Hi Everybody,

Recently we are including mice as another animal species into our
studies. Lysis protocols for whole blood specimens or lysis of
erythrocytes in spleen fractions however seem to be much more cumbersome
than in specimens from human or monkeys. Seperation of cells by
gradients (ficoll) seem to be different than seen in human or monkey
specimens.

Eventually, we want to perform "tetramer-staining" on mice specimens.
However, the streptavidin conjugates which we tested so far have quite
some background staining (between 0.2 and 2%) on mice T cells (CD4+ T
cells have even a higher background than CD8+ T cells.)

Does anybody out there have some good suggestions for:
1) Lysis of mice erythrocytes
2) gradients like ficoll specific for mice T lymphocytes
3) streptavidin conjugations to PE that have a "very" low background
staining on mice T lymphocytes


Thanks for your help. Depending on the amount of responses I will put
out a summary on the web.

Joern E. Schmitz, M.D.
Division of Viral Pathogenesis
Beth Israel Deaconess Medical School
Harvard Medical School
Boston



This archive was generated by hypermail 2b29 : Wed Apr 03 2002 - 11:53:42 EST