CCK-8 was used for the proliferation ability of A549 cells after treated with the Exo [20 (link)], and then, the CCK-8 reagent was added to the cell medium and incubated at 37°C for 2 h. Then, the absorbance value of OD450 nm was detected at 0 h, 24 h, 48 h, and 72 h, and a microplate reader (Beyotime Biotechnology, China) was used. EdU experiment was used to visualize the DNA expression of A549 cells (Beyotime Biotechnology, China) and then observed under the fluorescence microscope.
Microplate reader
Manufactured by Beyotime
Sourced in China
The Microplate reader is a laboratory instrument designed to measure and analyze the optical density or fluorescence of samples in a microplate format. It is commonly used in various fields, such as biochemistry, cell biology, and drug discovery, to quantify the presence or activity of specific substances within samples.
Automatically generated - may contain errors
Lab products found in correlation
A549 cells, by Beyotime (1 mentions)
Cyquant cell proliferation assay kit, by Thermo Fisher Scientific (1 mentions)
Mtt solution, by Beyotime (1 mentions)
T sod assay kit, by Beyotime (1 mentions)
Mda elisa kit, by Abcam (1 mentions)
Cck 8 assay kit, by Beyotime (1 mentions)
Protein detection kit, by Beyotime (1 mentions)
Cell counting kit 8 cck 8, by Sangon (1 mentions)
10 protocols using microplate reader
1
Proliferation and DNA Expression of A549 Cells
2
Cell Proliferation Quantification Using CyQuant
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Cell number assay was performed using CyQuant cell proliferation assay kit (ThermoFisher Scientific, MA, USA) for assessment of cell proliferation. Cells (4 × 103) with the corresponding treatments were incubated in a 96-well plate. After 48 h, freshly prepared 10 μL detection solution fluorescent-DNA-binding dye mixture (provided by kit) was added to each well. Afterwards, cells were incubated for another 5 min in dark. The optical density value was measured at 450 nm with a microplate reader (Beyotime, Shanghai, China).
3
MTT Cell Proliferation Assay
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The SNU-1 cells (5 × 103) were inoculated into 96-well plates. Cell proliferative ability was detected using sterile MTT solution (Beyotime) after cell culture for 12, 24, 48, 72 h. Absorbance at 570 nm was detected by a microplate reader (Sunnyvale, CA, USA) [19 (link)].
4
NAC Modulation of Oxidative Stress in S. uberis Mammary Infection
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The mMECs were cultured overnight in 6-well plates and treated with NAC 1 h prior to S. uberis challenge (MOI 5:1). Total superoxide dismutase (T-SOD) and malondialdehyde (MDA) activity were measured at 6, 12 and 24 h post challenge. The T-SOD activity was measured using a T-SOD assay kit in a microplate reader (Beyotime Biotechnology), whereas MDA activity was evaluated with an MDA ELISA kit (Abcam, Shanghai, China). The same kits were used to determine the activities of T-SOD and MDA in murine mammary glands. Herein, tissues were collected, washed with PBS and stored at −20 °C. Then, tissues were transferred into a mortar with liquid nitrogen, ground to a powder, homogenized with 0.5% Triton X-100 and centrifuged at 14,000× g for 5 min at 4 °C to collect supernatants.
5
Cell Proliferation Assay with IL-1β and siRNA
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The proliferative ability of cells was examined by a CCK-8 assay kit (Beyotime biotechnology, China) after being treated with the IL-1β and siRNA transfection. Cells inoculated in 96-well plates were cultured for 3 days, and at indicated time point, 15-μL CCK-8 reaction solution was added to the cell culture for 2-h incubation at 37 °C. Then, the medium was removed, and the cells were dissolved in 150 μL DMSO for 15 min. A microplate reader (Beyotime biotechnology, China) was adopted for determining the absorbance (OD450) in each well.
6
Immune Response Monitoring Protocol
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After anesthesia, the abdominal cavity was exposed, and 5 mL of blood was collected from the abdominal aorta immediately. Blood samples were placed in a test tube containing EDTA (Ethylene Diamine Tetraacetic Acid) anticoagulant to count the total number of white blood cells. The blood samples were stained with Turk (a cell stain, mainly composed of acetic acid and crystal violet) solution (1:10 dilution), and the total white blood cells (Includes neutrophils, eosinophils, basophils, lymphocytes, and monocytes) were counted repeatedly in the hemocytometer (count/ml). We performed the WBC test (Blood was collected through the tail vein) 5 times during the experiment, 1 day before irradiation, 3 weeks, 6 weeks, 9 weeks, and 12 weeks. We have organized this part of the data in this paper and redrawn the WBC time-varying curve into the text to supplement the previous research results. According to morphological standards, 10 cells were counted under a light microscope for analysis.
A tissue lysis kit was used to extract proteins from the lung samples and all lung tissues were cleared of blood during the analysis. BCA (Bicinchoninic Acid) protein detection kit (P0012, Beyotime, China) was used to determine the protein concentration. Enzyme–linked immunosorbent assay (ELISA) kit (PC188, Beyotime, China) was used to quantify CRP levels (Microplate reader, BIO-RAD).
A tissue lysis kit was used to extract proteins from the lung samples and all lung tissues were cleared of blood during the analysis. BCA (Bicinchoninic Acid) protein detection kit (P0012, Beyotime, China) was used to determine the protein concentration. Enzyme–linked immunosorbent assay (ELISA) kit (PC188, Beyotime, China) was used to quantify CRP levels (Microplate reader, BIO-RAD).
7
Cytotoxicity Assessment of SFN
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To prepare SFN stock solution, 10 mM SFN was dissolved in DMSO. HepG2.2.15 cells were seeded into 96-well plates at a density of 2000 cells/well. The effect of SFN was assessed at concentrations of 0, 0.1, 0.5, 1, 2, 5, and 10 μM. Cell viability at 48 h was determined by the CCK-8 assay. The absorbance was measured at 450 nm with a microplate reader (Beyotime, Shanghai, China).
8
Cell Viability Assay with CCK-8
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Cell Counting Kit-8 (CCK-8; Sangon) was used for the examination of cell viability of OC cells. At 0 h, 24 h, 48 h and 72 h after transfection, CCK-8 solution was pipetted to OC cells in the 96-well plates with 10 μL/well. Incubating for 2 h, the optical density (OD) value at 450 nm was detected with a microplate reader (Beyotime, Shanghai, China).
9
Curcumin Cytotoxicity Screening
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Cells cultured in 96-well plates were treated with different concentrations of curcumin for 0, 24 and 48 hours. CCK-8 reagent was added to the medium (1:100), and the cells were incubated for another 4 hours. Cell proliferation was assessed by measuring the optical density (OD) at 450 nm using a microplate reader (Beyotime, China) according to the manufacturer’s instructions.
10
Assessing Islet Antioxidant Defenses
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Islets were isolated post burn or sham treatment. Also, the activities of Mn-SOD were assessed spectrophotometrically by Microplate Reader using commercial kits according to the manufacturer's instructions (Beyotime, China).
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