The Edu assay was performed using the BeyoClick™ EdU-488 assay kit (Beyotime, C0071S) according to the manufacturer’s instructions. Briefly, 48 h after the EPA or DHA (50 or 100 μM) treatment, cells were exposed to 10 μM Edu for 2 h. Next, cells were fixed with 4% paraformaldehyde and permeabilized with 0.5% triton X-100. Subsequently, cells were incubated in the Click Additive Solution for 30 min and stained with DAPI. Fluorescence imaging of Edu+ cells was obtained with an Olympus Fluoview FV10C-O3 confocal microscope. The cells were further analyzed by calculating the ratio of Edu+ cells to the total number of cells.
Beyoclick edu 488 assay kit
Manufactured by Beyotime
The BeyoClick™ EdU-488 assay kit is a tool for detecting and quantifying cell proliferation. It utilizes a fluorescent dye to label and visualize cells undergoing DNA synthesis, a hallmark of cell division. The kit provides a straightforward method for measuring this process in a variety of cell types.
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Lab products found in correlation
Sp8 laser scanning confocal microscope, by Leica (1 mentions)
Celltiter 96 aqueous one solution cell proliferation assay mts kit, by Promega (1 mentions)
Enhanced immunostaining permeabilization buffer, by Beyotime (1 mentions)
Bovine serum albumin (bsa), by Solarbio (1 mentions)
Fluorescent microscope, by Nikon (1 mentions)
Cell counting kit 8 cck, by Beyotime (1 mentions)
Mts assay kit, by Promega (1 mentions)
5 protocols using beyoclick edu 488 assay kit
1
EdU-Based Cell Proliferation Assay
2
Evaluating Cell Proliferation and Viability
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Cells were maintained in 96-well plates (5 × 103 cells/well) for 24 h and transfected with siRNA. Cell viability and growth rate were assessed using CellTiter 96 AQueous One Solution Cell Proliferation Assay (MTS) kit (Promega, Madison, WI, USA) following the manufacturer’s instructions. Cell proliferation was assessed using the BeyoClick™ EdU-488 assay kit (Beyotime). Briefly, 3 × 104 cells were seeded in 24-well cover-glass bottom plates, incubated for 24 h, and transfected with siRNA. Then the cells were treated with 10 μM Edu for 2 h, fixed, permeabilized, and stained with the Click Additive Solution. Cell nuclei were stained with DAPI. Images were captured using a Leica SP8 laser-scanning confocal microscope. The cell proliferation rate was calculated as a percentage of EdU-positive cells per DAPI-positive cells. For the colony formation assay, cells were seeded into 12-well plates and treated with siRNA. After 14 days, visibly stained colonies were counted using Image J software.
3
Cell Proliferation Assay using EdU-488
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Cell proliferation was determined using BeyoClick™ EdU-488 Assay kit (Beyotime). mBMSCs were seeded into 6-well plates and incubated overnight. EdU working solution (10 µM) was added to the medium and incubated with cells for 3 h at 37 °C. Subsequently, 4% paraformaldehyde was used to fix cells for 15 min and washed off using PBS containing 3% bovine serum albumin (Solarbio). Furthermore, the mBMSCs were permeabilized with enhanced immunostaining permeabilization buffer (Beyotime) for 15 min. Then, the EdU detection was performed according to the manufacturer’s instructions. The nuclei were stained with Hoechst 33,342 for 10 min in the dark at room temperature. Fluorescence images were obtained by Olympus microscope (Olympus, Tokyo, Japan).
4
Proliferation Assays for Colorectal Cancer Cells
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A CCK-8 Cell Counting Kit (Beyotime, C0037) was used to assess the proliferation of HCT116 and SW480 cells. Cells were seeded in 96-well plates at a density of 2×104 cells/mL (100 µL/well) and incubated at 37 °C with 5% CO2. The optical density of the solution (OD450) in each well was measured by spectrophotometry (BioTek) on days 0, 1, 2 and 3 for HCT116 and SW480 cell, following addition of 10 µL CCK-8 reagent.
For EdU (5-ethynyl-2'-deoxyuridine) assay, cells were seeded in 96-well plates at a density of 5 × 104 cells per well in a 24-well plates and incubated at 37 °C with 5% CO2 overnight. The EdU incorporation assay was carried out according to the instructions for the BeyoClick™ EdU-488 assay kit (Beyotime, C0071S). After incubation with 10 μM EdU for 2 h, cells were fixed with 4% paraformaldehyde, permeabilized with 0.3% Triton X-100, and stained with fluorescent dyes (green). 1 × Hoechst 33342 was used to stain the cell nuclei (blue; 1:1000) at room temperature for 10 min. Cells were observed under a fluorescent microscope (Nikon).
For EdU (5-ethynyl-2'-deoxyuridine) assay, cells were seeded in 96-well plates at a density of 5 × 104 cells per well in a 24-well plates and incubated at 37 °C with 5% CO2 overnight. The EdU incorporation assay was carried out according to the instructions for the BeyoClick™ EdU-488 assay kit (Beyotime, C0071S). After incubation with 10 μM EdU for 2 h, cells were fixed with 4% paraformaldehyde, permeabilized with 0.3% Triton X-100, and stained with fluorescent dyes (green). 1 × Hoechst 33342 was used to stain the cell nuclei (blue; 1:1000) at room temperature for 10 min. Cells were observed under a fluorescent microscope (Nikon).
5
Cell Viability and Proliferation Assays
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The cell viability and growth rate were assessed at 48, 72 and 96 hours after transfection with siRNA, using MTS Assay Kits (Promega) according to the manufacturer's instructions. A standard immuno uorescence assay protocol was performed using the BeyoClick™ EdU-488 assay kit (Beyotime) following the manufacturer's instructions. In brief, following incubation with 10 μM Edu for 2 h, the indicated cells were xed, permeabilized and stained with the Click Additive Solution and DAPI. The images were acquired using a confocal laser-scanning microscope. The density of EdU-positive nuclei was calculated for cell proliferation analysis.
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