Varioskan lux multi function microplate reader

Viability of ALL cells was assessed using the CCK-8 assay. Briefly, NALM-6 and Jurkat cells (2 × 10⁴ cells/well) were cultured in 96-well plates with 0, 5, 10, 50, 100, and 150 μmol/L deferoxamine (DFO) (Amadis Chemical Company Limited, Shanghai, China) and incubated for 48 h at 37°C and 5% CO₂. Then, 10 μL of CCK-8 (SevenBio, CA, USA) was aseptically added and incubated for another 4 h. Subsequently, the optical density (OD) values were calculated at the absorbance of 450 nm via a Varioskan™ LUX multifunction microplate reader (Thermo Fisher Scientific, Waltham, MA, USA).
For detecting antiproliferative effects of DFO, ALL cells (3 × 10³ cells/well) were cultured for the indicated times (0, 1, 2, and 3 days). After incubation, the CCK-8 assay was performed with standard procedures.

HRECs with a density of 5 × 10³ cells per well were inoculated in a 96-well cell culture plate and incubated overnight at 37°C with 5% CO₂. Following the treatment with different concentrations of drugs for 24 h, 10 µl CCK-8 solution (Vazyme, cat. No A311-01/02) was added to each well and the cells were incubated at 37°C with 5% CO₂ for 4 h. Absorbance was measured at 450 nm using a Varioskan™ LUX Multi-function microplate reader (Thermo Fisher Scientific, Inc.).

The AZ samples obtained with different deamidation reaction times were tested using the aminase bioassay test kit (MAK310). All the samples were diluted with 80% ethanol to a protein concentration of 4 mg/mL. Moreover, the probes were added quantitatively and left for 2 h. The absorbance at λ_max = 570 nm was recorded (Varioskan LUX Multi-function microplate reader, Thermo Fisher Scientific, Waltham, MA, USA). DD was calculated according to the following Formula (1): $$DD\%=\frac{I_D-I_N}{I_S-I_N}\times 100\%$$
where I_D is the fluorescence intensity of the AZ dispersion; I_N is the fluorescence intensity of the untreated fresh zein dispersion; I_s is the fluorescence intensity of the zein dispersion treated with 3N sulfuric acid.