Human rhabdomyosarcoma (RD; CCL-136; ATCC) cells were maintained in Dulbecco’s modified Eagle Medium (DMEM) containing 10% fetal bovine serum (FBS; Life Technologies) at 37°C in a 5% CO2 incubator. Echovirus 11 strain (NCBI Accession No. OP764694) was isolated from a feces sample of a 24-day-old female neonate with enterovirus infection after passaging in the RD cells. RD cells were infected with echovirus at various multiplicity of infection (MOI) for 1.5 h in serum-free DMEM. The cells were washed with phosphate-buffered saline (PBS) and cultured in a completely fresh medium for various times as indicated until they were harvested. For autophagy induction experiments, cells were infected or mock-infected with echovirus for 1.5 h, then cultured in a complete medium containing rapamycin (Selleck, AY-22989) at indicated concentrations for the indicated times. For autophagy inhibition experiments, cells were cultured in DMEM containing indicated concentrations of 3-methyladenine (3-MA) (Selleck, S2767) for 2 h, followed by echovirus infection for 1.5 h, and then incubated with fresh DMEM for 16 h. Cell counting kit-8 (CCK8) (Vazyme, A311-01) assay was performed to examine the cytotoxicity of rapamycin or 3-MA to RD cells.
A311 01
Manufactured by Vazyme
Sourced in China
The A311-01 is a laboratory equipment product. It is a device designed for use in scientific research and analysis. The core function of the A311-01 is to perform specific laboratory tasks. Further details on the intended use or capabilities of the product are not available.
Automatically generated - may contain errors
Lab products found in correlation
A33978, by Thermo Fisher Scientific (9 mentions)
5 ethynyl 2 deoxyuridine edu assay, by RiboBio (2 mentions)
Crystal violet, by Solarbio (2 mentions)
Ccl 136, by American Type Culture Collection (1 mentions)
Ay 22989, by Selleck Chemicals (1 mentions)
S2767, by Selleck Chemicals (1 mentions)
Synergy ht microplate reader, by Agilent Technologies (1 mentions)
5 ethynyl 2 deoxyuridine (edu), by Beyotime (1 mentions)
15 protocols using a311 01
1
Echovirus 11 Infection in RD Cells
2
CCK8 Proliferation Assay Protocol
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CCK8 assay was used to evaluate cell proliferation according to the manufacturer's protocol (A311-01, Vazyme Biotech, Nanjing, China). U251 and T98G cells were seeded in 96-well-plates at a density of 5,000 cells/100 ul/well. Then, 10 ul of CCK8 solution was added to each well of the plates and incubated for 1 h at 37°C. Finally, the absorbance was analyzed at 450 nM by using a BioTek Synergy HT Microplate Reader.
3
Cell Viability Assay with CCK-8
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In 96-well plates, we seeded the cell suspension with 3×103 cells per well. The plate was then incubated for 2 hours at 37°C in the dark with 10 mL of CCK-8 labeling agent (A311-01, Vazyme) each well. The enzyme-labeled meter (A33978, Thermo) was used to measure the absorbance of the cells at 450 nm for 0, 24, 48, 72, and 96 hours in order to determine the vitality of the cells.
4
Cell Proliferation Assay Protocol
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Cells were seeded in 96-well plates (2000 cells/well), and cultured for the indicated time. Cell growth was measured by incubating cells with CCK-8 solution according to the manufacturer’s instructions (A311-01, Vazyme Biotech). The absorbance at 450 nm was then measured using a microplate reader. The experiment was done in triplicate.
5
Cell Viability Assay of U87 and U251 Cells
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Cell viability of U87 MG cells and U251 MG cells after transfection was detected by CCK-8. The cell suspension was inoculated at a density of 5 × 103 cells per well in a 96-well plate and incubated for 24 h. CCK-8 marker (10 μL; A311-01, Vazyme) was added to each well, and incubated away from light at 37°C for 2 h. Cell viability was assessed by detecting the absorbance of the enzyme marker (A33978, Thermo) at 450 nm on days 1, 2, 3, and 4, respectively. The average OD values were calculated and plotted on a line graph.
6
Cell Viability Assessment using CCK-8
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Cells were plated in 96-well plates at a density of 1 × 103 cells per well, following standard protocols (45 (link), 46 (link)). Following that, the plates were incubated in darkness at 37°C for 2 hours with CCK-8 labeling reagent (A311-01, Vazyme). The assessment of cell viability was carried out by measuring the absorbance at 450 nm using an enzyme-linked spectrophotometer (A33978, Thermo) at time intervals of 0, 24, 48, 72, and 96 hours.
7
Cell Viability Assay with CCK-8
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At first, 5×103 cells per well were seeded in the cell suspension in 96-well plates. All these cells were precultured. Following that, the plate was incubated with 10 mL of CCK-8 labeling solution (A311-01, Vazyme) in each well for 2 hours at 37° C in a dark environment. The enzyme-labeled meter (A33978, Thermo) was used to measure the cells’ absorbance at 450 nm in order to determine their health. Six measurements were made every 24 hours.
8
Cell Viability and Proliferation Assay
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The suspension of cells was seeded in 96-well plates at a density of 5×103 cells per well. After adding 10 μl of CCK-8 labeling agent (A311-01, Vazyme) to each well, the plate was incubated for 2 hours in the dark at 37°C. Cell viability was evaluated by measuring absorbance at 450 nm at 0, 24, 48, 72, and 96 hours using an enzyme-labeled meter (A33978, Thermo). The experiment was performed using a 96-well plate with 2×104 treated cells in each well, after the cells had adhered to the wall. The 5-Ethynyl-2’-deoxyuridine (EdU) assay was performed according to the manufacturer’s instructions (Ribobio, China), and cell proliferation was quantified using an inverted microscope.
9
Cell Proliferation Assay using CCK-8
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In this procedure, cells were plated in 96-well plates at a concentration of 3×10³ cells per well. Following cell seeding, 10 mL of CCK-8 solution (A311-01, Vazyme) was added to each well. The plates were then incubated in the dark at 37°C for 2 hours. The proliferation of cells was determined by measuring absorbance at 450 nm with a spectrophotometer (A33978, Thermo Fisher Scientific) at time points of 0, 24, 48, 72, and 96 hours.
10
Cell Viability Assay Protocol
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Each well of a 96-well plate was seeded with 2000 treated cells. Following this, the cells were subjected to treatment with the CCK-8 labeling reagent (A311-01, Vazyme) and evaluated at various time points, including days 1, 2, 3, 4, and 5.
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