I’m new to red blood cell-based assays. Anyone can help me answer the following questions? Thank you very much!
I plan to evaluate the effect of a compound on the hemolysis of human erythrocytes. I also want to assess whether or not this compound affects the GSH levels of erythrocytes. I will start with pure erythrocytes without preparing suspension from blood. I plan to treat the erythrocytes with the compound for at least 24-48 hours before the hemolysis and GSH assays.
1) If I plan to treat erythrocytes with the compound for 24-48 hours, what buffer (medium) can I use for this process? Saline? RPMI-1640 (10%FBS +P/S)? Any good recommendations to keep the best health status of the erythrocytes during the process?
2) I want to compare conditions with or without glucose supplementation for this process. Can I add glucose to the recommended buffer?
3) Since I will start with pure erythrocyte, what cell density (no. of erythrocyte/mL) should I start with?
4) I plan to run the assay in a 96-well format. Should I use a flat-bottom or round-bottom culture plate?
Thank you very much!
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