An EdU assay kit (Invitrogen; Thermo Fisher Scientific, Inc.) was used to determine MAC-T cell viability. Cells were plated into a 24-well plate (5x104 cells/well). Each well was supplemented with 50 µM EdU for 2 h at 37˚C. Subsequently, MAC-T cells were stained with DAPI (Beyotime Institute of Biotechnology) for 5 min in the dark at room temperature and then cultured with 1X ApolloR reaction cocktail supplied with EdU assay kit (Invitrogen; Thermo Fisher Scientific, Inc.) for 30 min at room temperature, cells were fixed with 4% paraformaldehyde for 15 min at room temperature. Cell viability was determined using a fluorescent microscope.
Edu assay kit
Manufactured by Thermo Fisher Scientific
Sourced in United States
The EdU assay kit is a laboratory tool used to detect and quantify cellular proliferation. It functions by incorporating a modified nucleoside, EdU (5-ethynyl-2'-deoxyuridine), into the DNA of actively dividing cells, which can then be visualized and analyzed using fluorescent detection methods.
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Lab products found in correlation
Cell proliferation reagent kit 1 mtt, by Roche (4 mentions)
Fluorescent microscope, by Olympus (4 mentions)
Image pro plus 6, by Media Cybernetics (3 mentions)
Microplate reader, by Thermo Fisher Scientific (2 mentions)
Hoechst 33342, by Thermo Fisher Scientific (2 mentions)
Hoechst 33342, by Merck Group (2 mentions)
Cell proliferation reagent kit 1, by Roche (2 mentions)
Cycletest plus dna reagent kit, by BD (2 mentions)
Edu assay, by Thermo Fisher Scientific (1 mentions)
Fluorescence microscopy, by Olympus (1 mentions)
Fluorescence microscopy, by Zeiss (1 mentions)
Iclick edu andy fluor 647 flow cytometry assay kit, by GeneCopoeia (1 mentions)
Methyl thiazolyl tetrazolium (mtt), by Merck Group (1 mentions)
Methyl thiazolyl tetrazolium (mtt), by Beyotime (1 mentions)
Cck 8 assay, by Dojindo Laboratories (1 mentions)
Exl800 microplate reader, by Agilent Technologies (1 mentions)
4 6 diamidino 2 phenylindole (dapi), by Beyotime (1 mentions)
Cell counting kit 8 cck 8 assay, by Dojindo Laboratories (1 mentions)
Elipse ti, by Nikon (1 mentions)
Dynabead, by Thermo Fisher Scientific (1 mentions)
30 protocols using edu assay kit
1
Evaluating MAC-T Cell Viability with EdU Assay
2
Cell Viability and Proliferation Assays
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Cell viability and proliferation were detected using cell counting kit 8 (CCK-8) and EdU assay. For CCK-8 assay, cells were seeded into 96-well plate and incubated for 24 h. CCK-8 reagent was added into culture medium for 2 h, and the optical density was measured at 450 nm using microplate reader (Thermo, USA). EdU assay was performed using EdU assay kit (Thermo, USA) in line with manufacturer's introduction.
3
Acute Cell Proliferation Post Needle Injury
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To determine the acute response to needle puncture injury, cell proliferation was assessed day 3 post injury (n = 7 per group) by EdU assay according to manufacturer’s instructions (ThermoFisher Scientific, Waltham, MA, USA). In brief: EdU (A10044, ThermoFisher Scientific, Waltham, MA, USA) was injected subcutaneously 24 h before sacrifice. Histological IVD-vertebrae sections were stained using an EdU assay kit (C10338, ThermoFisher Scientific, Waltham, MA, USA). One section per IVD was used for analysis. Proliferation was assessed 3 days after injury because this early timepoint should approximately correspond with the end of the initial acute inflammatory response to needle puncture, and the start of the proliferative phase of wound healing.
4
Cell Proliferation Analysis using EdU Assay
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Cell proliferation was also determined using the EdU assay kit (Thermo Fisher Scientific) according to the manufacturer’s instructions. Cells (1 × 105) were maintained in 6-well plates. After 48 h, 100 μL EdU was added for 2 h. Cells were incubated with 4% formaldehyde, followed by 0.3% Triton X-100 for 10 min. EdU-positive cells were analyzed using fluorescence microscopy (Olympus, Tokyo, Japan) at the 20× objective.
5
Cell Proliferation Measurement with EdU Assay
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Following the instructions from the manufacturer, an EdU assay kit (Thermo Fisher Scientific, Waltham, MA, USA) was used to measure proliferation of cells. The cells (2.0 × 104) were either visualized with fluorescence microscopy (Carl Zeiss Microscopy GmbH, Jena, Germany) or counted by flow cytometry (FACSCalibur) with an iClick EdU Andy Fluor 647 Flow Cytometry Assay Kit (Genecopoeia, Germantown, MD, USA). All experiments were performed in triplicate.
6
Cell Proliferation Assay with EdU
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A total of 5 × 104 cells/well was seeded into the 96-well plate. After 24 h, cells were cultured with DMEM media containing 5 μM Ethynyl deoxyuridine (EdU) (EdU assay kit, Thermo Fisher Scientific), 10% fetal bovine serum for 2 h. Then the cells were fixed with 4% formaldehyde at room temperature for 30 min and then treated with an Apollo reaction cocktail for 30 min. Finally, using Hoechst 33342 (Thermo Fisher Scientific) DNA staining and visualized under a fluorescent microscope.
7
Measuring Cell Proliferation with MTT and EdU
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Cell proliferation was measured by diphenyl tetrazolium bromide (MTT) assay and 5‐ethynyl‐2′‐deoxyuridine (EdU) detection with a MTT cell proliferation (Beyotime, China) and EdU assay kit (Invitrogen), respectively. Cells were seeded into 96‐well plates and transfected with miR‐133a, inhibitor or miR‐133a scramble, respectively. After 0, 24, 48, 72, and 120 h, cells were incubated with 10 μL of MTT (5 mg/mL, Sigma) for another 4 h at 37°C, followed by removal of the culture medium and addition of 150 μL dimethyl sulfoxide (DMSO). Absorbance values at a wavelength of 570 nm were recorded on a microplate reader.
After transfection, cells were exposed to 50 μmol/L of EdU for 4 h at 37°C and fixed in 4% paraformaldehyde for 10 min at room temperature. After being washed with a phosphate‐buffered saline (PBS, 140 mmol/L NaCl, 2.7 mmol/L KCl, 10 mmol/L Na2HPO4, and 1.8 mM KH2PO4), and permeabilized with 0.2% TritonX‐100 in PBS at 37°C for 30 min at room temperature. After being washed with PBS twice for 5 min, cells were reacted with 100 μL of 1 × Apollo reaction cocktail for 30 min. Then, the cells were stained with 100 μL of Hoechst 33342 (5 μg/mL) for 30 min and visualized under a fluorescent microscope.
After transfection, cells were exposed to 50 μmol/L of EdU for 4 h at 37°C and fixed in 4% paraformaldehyde for 10 min at room temperature. After being washed with a phosphate‐buffered saline (PBS, 140 mmol/L NaCl, 2.7 mmol/L KCl, 10 mmol/L Na2HPO4, and 1.8 mM KH2PO4), and permeabilized with 0.2% TritonX‐100 in PBS at 37°C for 30 min at room temperature. After being washed with PBS twice for 5 min, cells were reacted with 100 μL of 1 × Apollo reaction cocktail for 30 min. Then, the cells were stained with 100 μL of Hoechst 33342 (5 μg/mL) for 30 min and visualized under a fluorescent microscope.
8
Cell Proliferation and DNA Synthesis Assays
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After transfection, cell proliferation ability was evaluated by CCK-8 assay (Dojindo). Cells were cultured for 0, 24, 48 or 72 h in 96-well plates, after that, 10 μl of CCK-8 (5 mg/ml) was added to the culture medium in each well. The absorbance at 450 nm was measured by Exl 800 microplate reader (Bio-tek).
EdU assay was performed to determine DNA synthesis in proliferating cells by using an EdU assay kit (Invitrogen). After transfection, cells were cultured for 48 h, fixed with 4% paraformaldehyde and permeabilized by 0.3% Triton X-100. Then cells were incubated with 10 μM EdU for 2 h, and cell nuclei were stained with DAPI (5 μg/ml). The number of EdU-positive cells was counted under a microscope in five random fields (Olympum). All assays were independently performed in triplicate.
EdU assay was performed to determine DNA synthesis in proliferating cells by using an EdU assay kit (Invitrogen). After transfection, cells were cultured for 48 h, fixed with 4% paraformaldehyde and permeabilized by 0.3% Triton X-100. Then cells were incubated with 10 μM EdU for 2 h, and cell nuclei were stained with DAPI (5 μg/ml). The number of EdU-positive cells was counted under a microscope in five random fields (Olympum). All assays were independently performed in triplicate.
9
Quantifying Cell Proliferation by EdU Assay
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Cell proliferation was measured by EdU assay kit (Invitrogen). Cells were seeded into 96-well plates and transfected with Cav1.3 siRNA or GFP siRNA, respectively. After 48 h, cells were exposed to 50 µM of EdU for 4 h at 37°C and fixed in 4% paraformaldehyde for 10 min at room temperature. After being washed with a phosphate-buffered saline (PBS; 140 mM NaCl, 2.7 mM KCl, 10 mM Na2HPO4 and 1.8 mM KH2PO4) and permeabilized with 0.2% Triton X-100 in PBS at 37°C for 30 min at room temperature. After washing with PBS twice for 5 min, cells were reacted with 100 µl of 1X Apollo reaction cocktail for 30 min, and were stained with 100 µl of Hoechst 33342 (5 µg/ml) for 30 min. The nuclear was stained with 4′,6-diamidino-2-phenylindole (DAPI) (10 mg/ml) and visualized under a fluorescent microscope.
10
Cell Proliferation Assay for A549 and NCI-H460 Cells
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A549 or NCI-H460 cells (3×103 cells/well) were seeded in 96-well plates. After transfection with si-NC or si-LINc00887, cell proliferation was evaluated using a Cell Counting Kit-8 (CCK-8) assay (Dojindo Molecular Technologies, Inc.) according to the manufacturer's instructions. Cells were cultured for 0, 24, 48 or 96 h at 37°C and incubated with 10 µl CCK-8 reagent per well at 37°C for 1 h. The absorbance was measured at 450 nm using an Exl 800 microplate reader (BioTek China). The 5-ethynyl-2-deoxyuridine (EdU) staining assay was performed to determine DNA synthesis in proliferating cells using an EdU assay kit (Invitrogen; Thermo Fisher Scientific, Inc.). After transfection with si-NC or si-LINC00887, A549 or NCI-H460 cells were cultured at 37°C for 48 h, fixed with 4% paraformaldehyde at room temperature for 10 min and permeabilized with 0.3% Triton X-100 at room temperature for 10 min. Subsequently, cells were incubated with 10 µM EdU for 2 h at 37°C, and cell nuclei were stained with DAPI (5 µg/ml) at room temperature for 10 min. The number of EdU-positive cells was counted under a light microscope in five random fields (magnification, ×100; Olympus Corporation). All assays were independently performed in triplicate.
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