The MDA-MB-231, Hs578T and RAW264.7 cells (5×103)/well were seeded and incubated in 96-well plates at 37°C for 24 h. Conditioned medium (CM) or drugs (IL-13, 30 ng/ml; and/or αPD-L1, 15 µg/ml) were added and the cells were further incubated at 37°C for 0, 24, 48 and 72 h. Subsequently, 100 µl MTT reagent (1 mg/ml; Beyotime Institute of Biotechnology) were added to each well and the cells were incubated at 37°C for 4 h under the same conditions. Subsequently, 100 µl DMSO (Beijing Solarbio Science & Technology) were added to each well and the optical density at 490 nm was assessed using a full-wavelength multifunctional microplate reader (Tecan, Inc.). At least five wells/group were analyzed and the experiment was repeated three times.
Full wavelength multifunctional microplate reader
Manufactured by Tecan
Sourced in Switzerland
The Full-wavelength multifunctional microplate reader is a laboratory instrument designed to measure the optical properties of samples in microplates. It is capable of detecting a wide range of wavelengths, allowing for versatile applications in various fields of research and analysis.
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Lab products found in correlation
2 protocols using full wavelength multifunctional microplate reader
1
Cell Viability Assay with Cytokines
2
Assessing Peptide-Induced ROS in Bacteria
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The level of ROS induced by peptides in bacteria was assessed using the methods outlined by wang et al. [32 (link)]. E. coli CMCC44102 and S. aureus CMCC26003 were cultured to the exponential phase in MHB broth, washed thrice with PBS (pH 7.4), and subsequently diluted to a concentration of 5 × 108 CFU/mL. The bacterial suspension was treated with 2′,7′-dichlorofluorescein diacetate (DCFH-DA, Beyotime, Nantong, China) at a concentration of 10 µM and incubated at 37 °C for 30 min for fluorescence-probing purposes. Then, 190 µL of probe-labeled bacterial suspension was added to a 96-well plate along with 10 µL peptide samples. After incubation at 37 °C for another 30 min, fluorescence was measured using a full-wavelength multifunctional microplate reader (Tecan, Switzerland) with an excitation wavelength of 488 nm and emission wavelength of 525 nm. Three parallel tests were performed, and Mean ± SEM values were calculated. Polymyxin B was used as a positive control.
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