A colorimetric assay was used to analyze the cell viability of H3122 and H2228 cells using the CCK-8 reagent (Beyotime Biotechnology, Shanghai, China), respectively. Cells transfected with pcDNA-lincROR or pcDNA control were set as the experimental or the control group. After 24 h transfection, the cells were planted into 96-well plates supplemented with 10 μL of CCK-8 reagent and then incubated for 1 h at 37°C. Subsequently, the transfected cells were separately treated with crizotinib (TargetMol, Shanghai, China) at various concentrations (0, 0.25, 0.5, 1.0, and 5.0 μM) and then cultured for 24 h. Finally, the cell viability was analyzed by measuring the OD at 450 nm wavelength using the Tecan M200 multimode microplate reader (Tecan, Mechelen, Belgium).
M200 multimode microplate reader
Manufactured by Tecan
Sourced in Belgium, Austria
The M200 is a multimode microplate reader that provides optical detection across multiple measurement modes, including absorbance, fluorescence, and luminescence. It is designed to perform a variety of assays and can be used for diverse applications in life science research and drug discovery.
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Lab products found in correlation
3 protocols using m200 multimode microplate reader
1
Colorimetric Assay for Cell Viability
2
Red Blood Cell Hemolysis Assay
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Sterile 0.1 mM
PBS, pH 6, was prepared by addition of potassium dihydrogen phosphate
to sterile Dulbecco’s PBS and sterilized through a sterile
vacuum filter with a 0.22 μm pore size. Red blood cells (200
μL) were added to 1800 μL of unmodified starch, diaminated
starch, and chitosan having been dissolved/dispersed in 0.1 M sterile
Dulbecco’s PBS at pH 6 and 7.4 in a final concentration of
0.1, 0.2, 0.5, 1, 2, 3, and 5 mg/mL. Sterile 0.1 M PBS, pH 6 and 7.4,
was used as a negative control, and 1% (v/v) Triton X-100 solution
was used as a positive control. The mixtures were incubated at 300
rpm and 37 °C for 4 h and then mixed by inversion every 15 min.
At predetermined time points, mixtures were centrifuged at 13,400
rpm at 4 °C for 5 min, and the absorbance of the supernatant
was measured using a M200 multimode microplate reader (Tecan, Grödig,
Austria).18 (link) Each concentration was evaluated
in triplicate, and the percentage of hemolysis was calculated according
to the following equation(eq 5 ):
PBS, pH 6, was prepared by addition of potassium dihydrogen phosphate
to sterile Dulbecco’s PBS and sterilized through a sterile
vacuum filter with a 0.22 μm pore size. Red blood cells (200
μL) were added to 1800 μL of unmodified starch, diaminated
starch, and chitosan having been dissolved/dispersed in 0.1 M sterile
Dulbecco’s PBS at pH 6 and 7.4 in a final concentration of
0.1, 0.2, 0.5, 1, 2, 3, and 5 mg/mL. Sterile 0.1 M PBS, pH 6 and 7.4,
was used as a negative control, and 1% (v/v) Triton X-100 solution
was used as a positive control. The mixtures were incubated at 300
rpm and 37 °C for 4 h and then mixed by inversion every 15 min.
At predetermined time points, mixtures were centrifuged at 13,400
rpm at 4 °C for 5 min, and the absorbance of the supernatant
was measured using a M200 multimode microplate reader (Tecan, Grödig,
Austria).18 (link) Each concentration was evaluated
in triplicate, and the percentage of hemolysis was calculated according
to the following equation
3
Quantification of β-CD Modification on Rluc
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The amount of β-CD modified on a Rluc was determined by a sulfuric acid-phenol assay with slight modification [18 (link)]. The detailed results can be found from the supporting documents. In a typical test, a 30 μL of ice-colded glucose standard samples or β-CD-Rluc solution was mixed with 100 μL of concentrated sulfuric acid in microplate wells. 20 μL of aqueous 5% phenol solution was then added into the well. The plate was floated uncovered on near boiling (>90 °C) water bath for 5 min for color development, followed by cooling on ice for another 5 min. A microplate reader (Tecan M200 multimode microplate reader) was used to collect the absorbance of each well at 490 nm. The concentration of standard solution was plotted with the absorbance of each solution. The molar concentration of β-CD was thus calculated. The number of β-CD per protein was obtained by dividing the molar concentration of β-CD over the molar concentration of protein Rluc.
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