I am attempting to quantify a specific progenitor cell from sickle cell blood by flow cytometry. My cell yield was poor using ficoll separation and now I am trying red cell lysis. I have tried using Qiagen red cell lysis buffer (1X) for 10min at RT without good results. Thanks!
Try to wash after Lyse , and befor Staining with monoclonal antibodies.
And try to cahnge your lyse , according to the flowcytometry machine
Hi Alicia, what do you mean by a "specific progenitor cell from sickle cell blood"?
Sickle cell anemia is an inherited recessive mutation of hemoglobin.
We are trying to isolate endothelial progenitor cells- a marker for vascular repair. Sickle red cells are more resistant to osmotic lysis, thought to be due to cell damage from sickled state. Therefore, normal protocol for red cell lysis do not seem to be working on these cells. Thanks
Alicia, did you ever find an alternative lysing solution for the sickle blood?
Dear Alicia have you tried to lyse red cells by freeze thaw cycles in .01% saponin in PBS (1x).
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