I am performing hemolysis assay for antimicrobials but there's difficulty in selecting the concentrations of the antimicrobials. I have read the papers and MHC 90 was determined. Should I consider IC50?
Also I am facing problem with blood that I can use it on the same day. Next day might be due to oxidation RBCs sediment.
Can I store it in PBS?
Test a range of antimicrobial concentrations as a set of serial dilutions, to measure the hemolytic potency, instead of a single concentration.
Adam B Shapiro I am considering to test in form of serial dilutions but unable to identify which concentrations should I use.
I would use a high concentration based on the expected maximal blood level of the antibiotic when used as a drug, because any amount of hemolysis would be a serious side effect. Antibiotics are usually given at large doses. Maximal blood levels can be high such as tens of micromolar at the peak, although some of the drug is bound up by blood proteins such as albumin. My suggestion is to choose the highest concentration at which the substance is soluble in the assay medium as the top concentration. If you can get it to 64 micrograms/mL, that would be a reasonable starting point.