Cells were cultured in 96-well plate for 24 h, and then treated with echinatin (TargetMol, Boston, MA, USA) at different concentrations. The cell viability was determined by Cell Counting Kit-8 (CCK-8; Dojindo Molecular Technologies Inc., Rockville, MD, USA) according to the manufacturer’s guidelines, and the absorbance was quantified by an automated microplate spectrophotometer (BioTek Instruments Winooski, VT, USA) at 450 nm22 (link).
Automated microplate spectrophotometer
Manufactured by Agilent Technologies
Sourced in United States
The Automated Microplate Spectrophotometer is a lab equipment designed to measure the absorbance of samples in microplate format. It can quantify the concentration of various molecules, such as proteins, nucleic acids, and small molecules, by detecting their optical density across a range of wavelengths.
Automatically generated - may contain errors
Lab products found in correlation
Wst 1 assay, by Beyotime (4 mentions)
Wst 1 cell proliferation and cytotoxicity assay kit, by Beyotime (2 mentions)
Cell counting kit 8 cck 8, by Dojindo Laboratories (1 mentions)
Synergy ht, by Tecan (1 mentions)
Fitc dextran, by Targetmol (1 mentions)
Cck 8, by Beyotime (1 mentions)
Wst 1, by Agilent Technologies (1 mentions)
Wst 1, by Beyotime (1 mentions)
10 protocols using automated microplate spectrophotometer
1
Cell Viability Assay with Echinatin
2
Cytotoxicity Evaluation of ESI
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The cytotoxic activity of ESI was measured using the WST-1 assay (Beyotime, Jiangsu, China). A549 and H1299 cells were treated with ESI at various concentrations for different time points, and then washed once and incubated with WST-1 at 37 °C for 2 h. The plates were read on an automated microplate spectrophotometer (BioTek Instruments, Winooski, VT, USA) at 450 nm.
3
Cell Viability Determination via MTT Assay
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Cell viability was determined as described previously 25 (link). Briefly, cells were plated in 96-well plates at a density of 1000 cells per well, and 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide (MTT) was added at the end of experiment. The absorbance was measured at a wavelength of 570 nm on an automated microplate spectrophotometer (BioTek Instruments, Winooski, VT, USA).
4
Cell Viability Measurement using WST-1
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The WST-1 cell proliferation and cytotoxicity assay kit (Beyotime Biotechnology) was used to measure the cell viability according to the manufacturer’s instruction. In brief, the cells were seeded into the 96-well plate for indicated treatment, and then added with WST-1 for incubation 3 h at 37 °C. An automated microplate spectrophotometer (BioTek Instruments, Winooski, VT, USA) was used to measure the absorbance at 450 nm.
5
Cytotoxicity and Cellular Uptake of NAP
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WST-1 assay (Beyotime) was used to determine the cytotoxicity of NAP. In brief, A549 cells (2 × 103) were seeded in 96-well plate and treated with increasing concentration of NAP (up to 400 μM) for 48 h. WST-1 was added and the plate was read after 1.5 h on an automated microplate spectrophotometer (BioTek Instruments, Vermont, USA) at 450 nm according to the manufacturer’s instructions.
Colony-formation assay was performed to detect long-term survival of A549 cells in NAP treatment. 400 cells seeded in 12-well plates were treated with indicated concentration of NAP for 15 days. The plates were washed twice with PBS and fixed with methanol for 10 min at room temperature and then stained with 1% crystal violet for 5 min. The number of colonies was counted using ImageJ software (version 1.44I).
Cellular uptake assay was performed by using FITC-Dextran (TargetMol). A549 cells were seeded onto 12-well plate and incubated with FITC-Dextran (0.1 mg/mL) for 24 h, then the cells were treated with 50, 100, 150 and 200 μM of NAP for 24 h and 48 h, respectively. The fluorescence (excited at 488 nm and emitted at 530 nm) was measured with a multifunctional microplate reader Spark 10 M (Tecan, Synergy-HT).
Colony-formation assay was performed to detect long-term survival of A549 cells in NAP treatment. 400 cells seeded in 12-well plates were treated with indicated concentration of NAP for 15 days. The plates were washed twice with PBS and fixed with methanol for 10 min at room temperature and then stained with 1% crystal violet for 5 min. The number of colonies was counted using ImageJ software (version 1.44I).
Cellular uptake assay was performed by using FITC-Dextran (TargetMol). A549 cells were seeded onto 12-well plate and incubated with FITC-Dextran (0.1 mg/mL) for 24 h, then the cells were treated with 50, 100, 150 and 200 μM of NAP for 24 h and 48 h, respectively. The fluorescence (excited at 488 nm and emitted at 530 nm) was measured with a multifunctional microplate reader Spark 10 M (Tecan, Synergy-HT).
6
Proliferation Assay for TFEB Variants
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For the proliferation assay, CRC cells stably expressing TFEBWT or TFEBS142A/S138A were seeded in 96-well microplates at a density of 1.5 × 103 cells per well and were cultured for four days. One day after seeding, cells were stained with CCK-8 (Beyotime, Jiangsu, China). The automated microplate spectrophotometer (BioTek Instruments, Winooski, VT, USA) was employed to read the plate after 1.5 h at the 450 nm wavelength. Absorbance was normalized against the absorbance on the first day and was calculated. Each experiment was performed in triplicate.
7
Cell Viability Assay of CRC Cells
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The cell viability was determined by WST-1 (Beyotime). CRC cells were inoculated in 96-well plates (3 × 103 cells/well in 100 μL) and treated with 0, 0.8, 3.2 and 12.8 μM of INZ for 48 and 72 h, respectively. The WST-1 reagent was added to each well and incubated at 37 °C for 1.5 h, then the absorbance was measured at 450 nm using an automated microplate spectrophotometer (BioTek Instruments, Vermont, USA).
8
CRC Cell Viability Assay by WST-1
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The CRC cell viability was measured by WST-1 Cell Proliferation and Cytotoxicity Assay Kit (Beyotime Biotechnology, Shanghai, China), as previously described 22 (link). WST-1 was added, and cells were incubated for 2 h at 37°C, and the absorbance at 450 nm was measured on an automated microplate spectrophotometer (BioTek Instruments, Winooski, VT).
9
Lomitapide Cell Viability Assay
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Cells were exposed to lomitapide at indicated concentrations for 24 h, 48 h, and 72 h. The cell viabilities were assessed by WST-1 assay (Beyotime Biotechnology, Shanghai, China) according to the manufacturer's instructions [22] . The results were obtained by measuring the absorbance with an automated microplate spectrophotometer (BioTek Instruments, Winooski, VT, USA) at 490 nm.
10
Lomitapide Cell Viability Assay
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Cells were exposed to lomitapide at indicated concentrations for 24 h, 48 h, and 72 h. The cell viabilities were assessed by WST-1 assay (Beyotime Biotechnology, Shanghai, China) according to the manufacturer's instructions [22] . The results were obtained by measuring the absorbance with an automated microplate spectrophotometer (BioTek Instruments, Winooski, VT, USA) at 490 nm.
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