To all of you who use ammonium chloride lysing of RBC in whole blood samples from mice for surface markers: I need some advice concerning lysing of murine RBC. I have just started to work with whole blood from BALB/c mice, and are trying to lyse the RBC with a NH4Cl-solution. My problem is that my samples aren't completely lysed, and one can see that the pellets are redish in some cases. My qustions are: What is the optimal g-value for centrifugation of the samples to get maximum lysis? We use 1200rpm (270g) for 10 min. Is there anything else we should be aware of in using NH4Cl lysing - both concerning the lysing method and analyzing the samples on the flowcytometer? So if anyone have some answers/advices or best of all a detailed protocol of how to lyse RBC in murine whole blood they could send me, I will be extremly happy! Thanks in advance! Anita Haugan Department of Vaccinology National Institute of Public Health, Norway
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