Mouse RBC fix/permeabilization protocolfor FACS

From: Dana Levasseur (Dana_Levasseur@microbio.uab.edu)
Date: Thu Aug 12 1999 - 18:04:25 EST


Hello Everyone,

I am trying to find a protocol for fixing and permeabilizing mouse
erythrocytes to allow intracellular staining for fetal hemoglobin levels
with a HbF-FITC antibody.  I can only assume that C57Bl/6 RBCs are
significantly more susceptible to hemolysis than human RBCs.  People have
treated human RBCs  for hours at room temp in 4% formaldehyde before
permeabilization with Triton.   Mouse RBCs lyse after short duration
exposure to 4% formaldehyde at 4 degrees and a single pelleting at low
speed.  Any ideas on how to handle such a fickle beast would be greatly
appreciated?!!

Dana 
Dana Levasseur
Dept. of Biochemistry and Medical Genetics
Bevill Biomedical Research Building, Rm 263
University of Alabama at Birmingham
Birmingham, Alabama  35205

e-mail:  dnl@uab.edu



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