The hemolytic test was carried out by using 2% erythrocyte suspension, which was prepared from human blood according to the previous description (Muñoz et al., 2006 (link)) with partial modifications. No hemolysis and 100% hemolysis were determined for controls with normal saline (NS) and 0.1% Triton X-100, respectively. The AMPs BP21 (The final concentration was 8, 16, 32, or 64 μmol L-1) was mixed with 2% red blood cells and incubated at 37°C for 1 h. After diluted 600 times, commercial Prochloraz was mixed with erythrocyte suspension. The samples were centrifuged at 1,000 ×g for 5 min, and the supernatant was transferred to 96-well plates. Release of hemoglobin was determined by OD540, and the data were measured by a Multiskan Spectrum microplate spectrophotometer. The hemolytic activity of peptide was calculated as the percentage of total hemoglobin released compared with that released by incubation with 0.1% Triton X-100.
Spectrum microplate spectrophotometer
Manufactured by MultiSciences Biotech
The Spectrum microplate spectrophotometer is a versatile instrument designed for absorbance measurements in microplates. It provides accurate and reliable data for a wide range of applications in life science research and development.
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Lab products found in correlation
2 protocols using spectrum microplate spectrophotometer
1
Hemolytic Activity of Antimicrobial Peptides
2
Glioblastoma Cell Viability Assay
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Influence of EA on the cell viability of human glioblastoma cell lines U251 MG was assessed by the cell counting kit-8 (CCK-8 kit), which was purchased from Dojindo China CO., Ltd (Beijing, China). Put simply, the cells were seeded in a 96well plate at a density of 0.8x10 4 cells/well when cells reached logarithmic growth, and incubated for 24 h until a 70% confluent layer was reached, the cells were exposed to various concentrations of EA (0-200µM) for 24 h, 48 h and 72 h respectively. At each time point, 100µl of fresh medium containing 10µl CCK-8 solution was added to each well and incubated at 37ºC for 0.5 h. The absorbance at 450 nm was measured on a Multiskan Spectrum Microplate Spectrophotometer. Each group was repeated in three wells.
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