Hi
I´m working with RBCC, and I´d like to assay viability and senescence of red and white cells. I was told to lyse red cells, but I don´t know how could I do it.
Also I wonder if I can stain with Anexin V, run cytometry for red cells, then lyse them, stain with 7-AAD and run cytometry for withe cells.
Thanks for your help
Regards,
Analía
Hi Analia,
You can use ACK lysis buffer (https://www.qualitybiological.com/product/ack-lysing-buffer/) to lyse the red blood cells, after spinning down your cells, aspirate the buffer and add 1ml ACK lysis buffer and incubate at RT for ~3.5min for splenocytes, 2min for bone marrow cells. For blood, you can either incubate at RT for 15min-20min followed by 3 times wash with FACS buffer or 5min RT first and wash and add another 1ml for another 5min.
To stain for RBCs, you can use TER-119 antibody (Catalog # 12-5921-82) together with your other surface staining antibodies, but I don't think you can still see them if you do RBC lysis after staining. All your white blood cells shouldn't express any TER-119, and all your RBCs will be brightly labeled TER-119+.
Hope this is helpful to you,
Zhixin
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