Defibrinated sheep blood (ThermoScientific, Oxoid Ltd., Hampshire, UK) was washed several times with sterile PBS (Sartorius) until the supernatant became clear. The blood cells were centrifuged at 1,700 g for 10 min between each wash. Approximately 109 erythrocytes were incubated with 1 ml of various concentrations of AA in PBS supplemented with 1% bovine serum albumin (BSA, VWR Chemicals, Solon, OH, USA) for 1 h at 37°C. The color intensity of the supernatant was determined at 405 nm in a Multiskan SkyHigh microplate reader (ThermoScientific, Life Technologies Holdings Pte Ltd., Singapore). Percentage hemolysis was calculated in comparison to ddw-exploded erythrocytes, which was set to 100% (Sæbø et al., 2023 (link)).
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