SH-SY5Y cells were plated in 96-well plates (10,000/well) for 48 hours, and 3-(4, 5-dimethylthiazol-2-yl)-2, 5-diphenyltetrazolium bromide (MTT, 5 mg/mL, 20 µL) was added to the cells and incubated for 4 hours at 37°C. The medium was removed, and dimethyl sulfoxide (150 µL) was added to the cells for 20 minutes at 37°C. The optical density (OD) values were measured with an automatic microplate reader (Bio-Tek, Winooski, VA, USA) at 492 nm. LDH activity was measured using LDH release assay kits, and the OD values were measured with an automatic microplate reader (Bio-Tek) at 450 nm.
Automatic microplate reader
Manufactured by Agilent Technologies
Sourced in United States, China
The Automatic Microplate Reader is a laboratory instrument designed to measure and analyze the absorbance, fluorescence, or luminescence of samples in a microplate format. It provides precise and automated data collection for various applications, such as enzyme-linked immunosorbent assays (ELISAs), cell-based assays, and screening experiments.
Automatically generated - may contain errors
Lab products found in correlation
Cck 8 assay, by Dojindo Laboratories (5 mentions)
Cell counting kit 8 assay, by Dojindo Laboratories (5 mentions)
Cell counting kit 8 cck 8, by Dojindo Laboratories (4 mentions)
Cck 8 solution, by Biosharp (4 mentions)
Cell counting kit 8 (cck8), by Dojindo Laboratories (3 mentions)
Cck 8, by Dojindo Laboratories (3 mentions)
Cck 8 solution, by Beyotime (2 mentions)
Cck 8 reagent, by Beyotime (2 mentions)
Cck 8 solution, by Dojindo Laboratories (2 mentions)
Cell counting kit 8 cck, by Dojindo Laboratories (2 mentions)
Methyl thiazolyl tetrazolium (mtt), by Merck Group (2 mentions)
Dcfh da, by Beyotime (2 mentions)
Phosphate buffered saline (pbs), by Merck Group (2 mentions)
Mtt solution, by Merck Group (2 mentions)
Prism 8, by GraphPad (2 mentions)
Mda kit, by Nanjing Jiancheng (1 mentions)
Sod kit, by Nanjing Jiancheng (1 mentions)
Crystal violet, by Beyotime (1 mentions)
Dimethyl sulfoxide (dmso), by Thermo Fisher Scientific (1 mentions)
Methyl thiazolyl tetrazolium (mtt), by Thermo Fisher Scientific (1 mentions)
107 protocols using automatic microplate reader
1
SH-SY5Y Cell Viability and Cytotoxicity Assay
2
Quantifying CRMP2 Protein Levels
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CRMP2 levels in the supernatant of disparate groups were detected using human CRMP2 ELISA kit (LifeSpan Bioscience, Seattle, WA, USA) according to the manufacturer’s instructions. The absorbance intensity of each sample was detected at 450 nm wavelength using an automatic microplate reader (BioTek Instruments, Winooski, VT, USA).
3
Cell Proliferation Assay for MKN-45/BGC-823
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The proliferation rate of MKN-45/BGC-823 cells was detected by the Cell Counting Kit-8 assay (Dojindo Laboratories, Kumamoto, Japan). Ten thousand cells were seeded in a 96-well plate, and 10 μL of CCK-8 solution were added to each well at the same time every day. After a 2-h incubation, the absorbance at 450 nm of the experimental wells was measured with an automatic microplate reader (BioTek, Winooski, VT, USA).
4
Liver Antioxidant Enzyme Quantification
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The MDA and SOD contents in liver tissues were detected using an MDA kit and a SOD kit (both kits were purchased from Nanjing Jiancheng Bioengineering Institute), respectively, according to the manufacturers’ instructions. The procedures are briefly described as follows:
SOD: The liver samples were mixed with the reagent and incubated at 37 °C for 20 min, and the absorbance at a wavelength of 450 nm was measured. The level of SOD activity and inhibition rate were calculated as follows: SOD activity (U/ml) = SOD inhibition rate/50% × dilution ratio in the reaction system/concentration of the sample protein to be tested; SOD inhibition rate = [(control A − blank control A) − (testing A − blank testing A)] × 100/(control A − blank control A).
MDA: The proteins were mixed with the reagent, incubated in a water bath at 95 °C for 40 min, cooled down in running water and centrifuged at 3500 rpm for 10 min. The MDA content was calculated as follows: MDA content (nmol/ml) = (absorbance of the testing tube − absorbance of the testing blank tube)/(the absorbance in the standard tube − the absorbance in the standard blank tube) × standard concentration (10 nmol/ml) × dilution ratio of the sample before the test. The absorbance values were obtained at a wavelength of 532 nm.
5
Cell Proliferation and Colony Formation Assay
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Cell proliferation was assessed using the Cell Counting Kit (CCK)-8 assay (Dojindo Laboratories, Kumamoto, Japan). Five thousand cells were added to each well of 96-well plates. 10 μl of CCK-8 solution was subsequently added to each well at four time points. After 2 h of incubation at 37 °C, the absorbance at 450 nM was measured using an automatic microplate reader (BioTek, Winooski, VT, USA). The experiments were repeated three times.
For the colony formation assays, cells were seeded in 6-well plates (300 cells per well) and incubated at 37 °C for 14 days. For colony scoring, the cells were fixed in 0.5 ml of methanol for 30 min and were stained with crystal violet (Beyotime Biotechnology, Nantong, China) for 15 min. The numbers of colonies were counted and expressed as means ± SEM of at least three independent experiments.
For the colony formation assays, cells were seeded in 6-well plates (300 cells per well) and incubated at 37 °C for 14 days. For colony scoring, the cells were fixed in 0.5 ml of methanol for 30 min and were stained with crystal violet (Beyotime Biotechnology, Nantong, China) for 15 min. The numbers of colonies were counted and expressed as means ± SEM of at least three independent experiments.
6
MTT Assay for GC Cell Viability
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The effects of CT on GC cell viability were measured using an MTT assay. Cells (1×104) were seeded in 96-well culture plates, and treated various concentrations (1, 3, 10, 30 and 100 μM) of CT or 5-FU for 24 h. And then, 15 μL of MTT solution (5 mg/mL) was added to each well and incubated at 37°C for 2 h, each well was then removed, and 100 μL DMSO was added to each well to dissolve formazan crystals. Absorption intensity was analyzed using an automatic microplate reader (BioTek Instruments Inc., Winooski, VT) at 490 nm, and results were used to calculate cell viability.
7
MTT Assay for Cell Viability
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Cells (1×103/well) were plated in 96-well plates and MTT (5 mg/ml; 20 µl; Thermo Fisher Scientific, Inc.) was added into each well for 4 h at 37°C. Dimethyl sulfoxide (150 µl; Thermo Fisher Scientific, Inc.) was subsequently added into the wells for 20 min at 37°C. Optical density values were measured with an automatic microplate reader (BioTek Instruments, Inc., Winooski, VT, USA) at 492 nm.
8
Cytotoxicity of Shikonin and Analogs
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Briefly, the human lung cancer A549, NCI-H23, and NCI-H460 cells and normal lung IMR-90, liver L-02, and stomach GES-1 cell lines were counted and seeded into 96-well culture plates followed by adherence for 24 h. Then, the cells were treated with different concentrations of shikonin, fluorouracil (5-FU), HEDMNQ, and HHDMNQ (0, 1, 3, 10, 30, and 100 μM) for 24 h. Followed by starvation for 2 h, 10 μL of CCK-8 solution (Beyotime Biotechnology, Beijing, China) was added to each well. After incubation for 1 h at 37°C, the absorbance of the solution was measured at 450 nm with an automatic microplate reader (BioTek Instruments, Inc., Winooski, VT).
In addition, the A549, NCI-H23, NCI-H460, IMR-90, L-02, and GES-1 cells were seeded in 96-well plates. According to the IC50 value of shikonin, 5-FU, HEDMNQ, and HHDMNQ in human lung cancer and normal cell lines, the cells were treated with HEDMNQ and HHDMNQ for 3, 6, 12, 24, and 36 h. Next, 10 μL of CCK-8 solution was added to the cells and measured by using an automatic microplate reader.
In addition, the A549, NCI-H23, NCI-H460, IMR-90, L-02, and GES-1 cells were seeded in 96-well plates. According to the IC50 value of shikonin, 5-FU, HEDMNQ, and HHDMNQ in human lung cancer and normal cell lines, the cells were treated with HEDMNQ and HHDMNQ for 3, 6, 12, 24, and 36 h. Next, 10 μL of CCK-8 solution was added to the cells and measured by using an automatic microplate reader.
9
OSCC Cell Co-culture with Neutrophils
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For the cell co-culture, 3 × 103 OSCC cells were seeded into each well of 96-well plates in the presence or absence of neutrophils (1:3 ratio). After 24 h, the neutrophils were removed from the co-culture system. Subsequently, methylthiazolyltetrazole (MTT) measurements were performed at different time points (0, 24, 48, 72, and 96 h). A total of 20 µl of MTT (5 mg/ml) was added into each well, and the cells were further incubated for 4 h. Then, the media were discarded, and 150 µl of DMSO was added. The plate was shaken at room temperature for 15 min, and absorbance was read at 490 nm on an automatic microplate reader (Bio-Tek Instruments, Winooski, VT, USA).
10
Sandwich ELISA for PD1ACR and PDL1CAR Binding
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A sandwich ELISA was performed to evaluate the binding ability of both PD1ACR and PDL1CAR to PD-L1, as described earlier.52 (link) In brief, 96-well plates were seeded with transduced 293T cells (PD1ACR, PDL1CAR, and mock). Untransduced 293T cells (blank) were used as a negative control. Each well was washed, and recombinant human PD-L1 Fc antigens were added (R&D Systems, Minneapolis, MN, USA) at different dilutions. Next, supernatants were collected and added to another 96-well plate, which was preliminarily coated with an anti-IgG (Fc) antibody, followed by the addition of a biotinylated IgG (Fc) detector antibody and a detection conjugate (Aviva Systems Biology Corporation, San Diego, CA, USA). After substrate detection, the optical density at 450 nm (OD450) was measured with an automatic microplate reader (BioTek, Winooski, VT, USA).
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