To detect the in vitro cytotoxicity of LPCE micelles, an MTT assay was used. Briefly, 293T cells were plated in 96-well plates (5 × 103 cells/well) overnight at 37 °C. When the number of cells was as required, different concentrations of LPCE were added to the cell plates. PEI25K was used as a standard control. After transfecting for 24 h at 37 °C, 20 μL of MTT solution was added to stop the reaction, and the cells were incubated for another four hours. Subsequently, the medium in the plate was replaced with 150 μL of dimethyl sulfoxide, and the plates were shaken on a shaker for 5 to 15 min. Finally, the absorbance of each hole was measured at 570 nm using a SpectraMax absorbance reader (Molecular Devices, San Jose, CA, USA).
Spectramax absorbance reader
Manufactured by Molecular Devices
Sourced in United States, China
The SpectraMax Absorbance Reader is a versatile laboratory instrument designed to measure the optical absorbance of samples. It is capable of performing rapid and accurate absorbance measurements across a wide range of wavelengths, enabling users to quantify the presence and concentration of various analytes in their samples.
Automatically generated - may contain errors
Lab products found in correlation
Methyl thiazolyl tetrazolium (mtt), by Yeasen (2 mentions)
Tnf α, by Boster Bio (1 mentions)
Il 1β, by Mlbio (1 mentions)
Dimethyl sulfoxide (dmso), by Solarbio (1 mentions)
Mtt assay kit, by Yeasen (1 mentions)
D1500, by R&D Systems (1 mentions)
Cck 8, by Elabscience (1 mentions)
Cck 8, by Molecular Devices (1 mentions)
Cck 8, by Dojindo Laboratories (1 mentions)
Cck 8 assay kit, by Vazyme (1 mentions)
Cck 8 assay, by Yeasen (1 mentions)
Cck 8 solution, by Dojindo Laboratories (1 mentions)
Cck 8 solution, by Molecular Devices (1 mentions)
Dp80 ccd camera, by Olympus (1 mentions)
Bx41 light microscope, by Olympus (1 mentions)
Novocyte flow cytometer, by Agilent Technologies (1 mentions)
Novoexpress 1, by Agilent Technologies (1 mentions)
Annexin 5 fitc pi apoptosis detection kit, by Vazyme (1 mentions)
Exfect transfection reagent, by Vazyme (1 mentions)
Mtt cell proliferation assay kit, by Beyotime (1 mentions)
32 protocols using spectramax absorbance reader
1
In Vitro Cytotoxicity Assessment of LPCE Micelles
2
Cell Viability Evaluation by MTT Assay
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After transfection for 24 h, the U-2 OS and 143B cells were seeded into 96-well plates at a density of 104 cells per well. An MTT assay kit (Cat: QF0025, Qiancheng Biotech, Shanghai, China) was used to evaluate cell viability at each time point according to the manufacturer’s instruction. The SpectraMax Absorbance Reader (Molecular Devices, San Francisco, CA, USA) was used to measure the absorbance at 490 nm. The data are represented in terms of mean ± s.d. for sextuple wells.
3
Fatty Acid Cytotoxicity in Liver Cells
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Fatty acid (including octanoic acid, decanoic acid, lauric acid, and oleic acid) were dissolved in PBS to form the stock solution at 8 mM and diluted by the serum‐free medium. For the viability assay, LO2 cells were seeded into 96‐well plates at a density of 1 × 104 cells per well and cultured overnight. Then, the cells were treated with different concentrations of FA (0, 50, 100, 200, 400, and 800 µM) for 24 hr. For further studies, LO2 cells were also treated with 20 mM AP and 600 µM RFP for 24 hr and then incubated with 200 µM FA for an additional 24 hr. Then, 10 µl of MTT solution was added to each well and incubated for 4 hr at 37°C. Next, the supernatant was carefully removed, the purple formazan crystals were dissolved in 150 µl DMSO, and the absorbance value at 490 nm was measured by a SpectraMax® absorbance reader (Molecular Devices).
4
Serum Cytokine Profile Analysis
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Serum was collected and cytokine levels were measured using an ELISA kit to determine the expression of alanine aminotransferase (ALT) (Servicebio), aspartate aminotransferase (AST) (Servicebio), PGE2 (Mlbio, Shanghai, China), TNF-α (Boster Biological Technology, Pleasanton, CA, USA), and IL-1β (Mlbio) according to the manufacturer’s instructions. After incubation, absorbance was finally read at 450 nm on a SpectraMax Absorbance Reader (Molecular Devices, Sunnyvale, CA, USA).
5
Pericyte Viability Evaluation with MTT
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The viability of pericytes was evaluated with MTT (Amersco Grade, Shanghai, China) according to the manufacturers’ protocols (Yu et al., 2019). Pericytes were seeded into 96-well plates at 5000 cells per well. After incubation, the supernatant was discarded and replaced with fresh culture medium. Thereafter, MTT, at a concentration of 5 mg/mL in PBS, was added to the medium, 20 μL per well, and incubated for 3 hours in the culture chamber. Next, dimethyl sulfoxide (Solarbio) was added at 150 μL per well for 10 minutes. Finally, optical absorbance was read at 562 nm using a SpectraMax Absorbance Reader (Molecular Devices, Silicon Valley, CA, USA).
6
MTT Assay for Cell Viability
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The viability of cells was explored via the MTT assay kit (CAT#40206ES76, Yeasen) as described by the vendor. Concisely, cells were planted and transfected in a 96-well plate. Cells were then stained with MTT at different time points (0 h, 24 h, 48 h, and 72 h), and the optical density of every sample at 490 nm was assessed using a SpectraMax Absorbance Reader (Molecular Devices, San Francisco, United States).
7
Cell Viability Assay with MDA-MB Cell Lines
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MDA-MB-231 and MDA-MB-157 cells were transiently transfected with the corresponding plasmids and siRNAs. The next day the transfected cells (3 × 103/well) were plated in 96-well plates in triplicates. Cell viability was assessed at 24, 48, 72 and 96 h, respectively. CCK-8 reagent (10 μL/well) was added to each well and incubated at 37 °C in dark for 2 h. The absorbance at 450 nm was examined with the SpectraMax Absorbance Reader (Molecular Devices, USA).
8
Quantifying Mouse IFNβ Levels
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Culture medium was collected from the cells. Using the mouse IFNβ ELISA kits (Bioswamp, Wuhan, China) according to the instructions, the OD values at 450 nm were determined by SpectraMax® Absorbance Reader (Molecular Devices Corporation, USA).
9
Quantification of IL-15 in HFRS and HTNV
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The levels of IL-15 in plasma samples of HFRS patients or in the culture supernatant of HUVECs infected with HTNV were detected using an ELISA kit (D1500, R&D, US) respectively according to manufacturer’s instructions. SpectraMax Absorbance Reader (Molecular Devices, US) was used to determine the absorbance at 450 nm.
10
Cell Viability and Colony Formation Assay
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To determine cell viability, cell counting kit‐8 (CCK‐8, Elabscience) was used. A total of 3 × 103 cells were seeded and cultured in 96‐well microplates using 100 μL of medium per well. After culturing for 24 h, 10 L of CCK‐8 reagent was added to each well, and the cells were cultured for 90 min. In all experiments, three replicates were performed. Wells without cells were used as blanks, and cell proliferation was measured by absorbance. Absorbance analysis was performed at 450 nm using a SpectraMax Absorbance Reader (Molecular Devices). In the colony‐forming assay, cells were seeded into six‐well plates at a density of 1 × 103 cells per well in DMEM with 10% FBS and cultured for 14 days to allow colony formation. Then, 4% polyformaldehyde was applied at room temperature to the plate after it was washed in cold PBS. In the next step, they were dyed at room temperature for 30 min with 1% crystal violet. Each experiment was performed three times, counting colonies over 100 cells by using ImageJ.
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