NH4Cl lysing of murine whole blood

From: Haugan, Anita (anita.haugan@folkehelsa.no)
Date: Thu Nov 16 2000 - 04:35:39 EST


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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