I want to use RBC lysing buffer to remove RBCs from my primary rat lung cell suspension prior to cell sorting (FACS). I am using the BD Pharm Lyse buffer- I'd appreciate any tips or advice!
Thank you for your sharing your protocol Denisa Nanushaj
What volumes/ ratios of cell suspension & buffer did you use for the splenocytes? I am working with lung cells so I will have to optimise the ratio for my samples
If you have pharm Lyse buffer go with it's recommended protocol, it works well. It comes in 10X potency, make 1X and use 1 mL for 100 uL sample (cells suspension). Vortex and incubate at 37 degree for 20-30 minutes, followed by centrifugation and washing.
Alternatively you may prepare ammonium chloride based lysis buffer in lab (we are using this for 2 years.
Thanks for your insight Jitendra Kumar Shandilya! I'll definitely check out their protocol.
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