I would like to do a FACs analysis but feel that there are too many RBCs. As I don't have very many samples, I would prefer to make a lysis buffer instead of ordering one.
is there a reason you need to lyse the RBCs as opposed to mechanically(centrifugation) separating them?
I centrifuged them several times but still have a lot of remaining RBCs and think there may be too many to just gate out.
Hi Elizabeth, it seems that most people are using commercial RBC lysis buffers. I found a publication that describes the composition of a RBC lysis buffer for flow analysis of mouse spleen:
http://iai.asm.org/content/79/2/674.full
Hope this helps.
Marc-André,
From what I understand mice RBCs are harder to lyse than human RBCs. I really appreciate the file but I think that particular solution would work best with human RBCs.
Fragility of red blood cells is discussed in detail in the very old article.
I used this technique personally with mouse blood sample. It's not expensive and you can try it by yourself and see if it's convenient for you.
Hello Elizabeth,
Use ACK lysis buffer to lyse RBC. It's highly effective. I'm using for long time to isolate WBC and it worked very smoothly. If you have any further question,let me be with you.
Best regards,
Babu
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