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
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