After culture in a 6-well plate, the NB cells were stained using the BeyoClickTM EdU‐488 kit (C0071S, Beyotime, China) as the instructions. Fluorescence microscopy was then used to examine the stained cells. The Image J software (National Institutes of Health) was employed to quantify the proportion of positive cell nuclei to the total number of cell nuclei.
Beyoclick edu 488 kit
Manufactured by Beyotime
Sourced in China
The BeyoClick™ EdU-488 Kit is a product designed for the detection of cell proliferation. The kit utilizes a modified nucleoside, EdU (5-ethynyl-2'-deoxyuridine), which is incorporated into the DNA of proliferating cells. The incorporated EdU is then detected using a fluorescent dye, allowing for the visualization and quantification of cell proliferation.
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Lab products found in correlation
Lsm 710, by Zeiss (8 mentions)
Microplate reader, by Bio-Rad (2 mentions)
Edu st067, by Beyotime (1 mentions)
Cell counting kit 8 (cck8), by Dojindo Laboratories (1 mentions)
Fluorescence microscope, by Olympus (1 mentions)
Transwell plate, by Corning (1 mentions)
Cck 8 kit, by GLPBIO (1 mentions)
Hoechst 33342, by Beyotime (1 mentions)
Dm 5000b fluorescence microscope, by Leica (1 mentions)
Hoechst, by Beyotime (1 mentions)
Inverted fluorescence microscope, by Zeiss (1 mentions)
Hoechst staining kit, by Beyotime (1 mentions)
Tubulin tracker deep red, by Thermo Fisher Scientific (1 mentions)
Sinps, by Merck Group (1 mentions)
Cpt1a, by Abcam (1 mentions)
4 6 diamidino 2 phenylindole (dapi), by Beyotime (1 mentions)
Gapdh, by Abcam (1 mentions)
Goat anti rabbit igg, by Abcam (1 mentions)
Caspase 3, by Abcam (1 mentions)
Fluorescein isothiocyanate (fitc), by Solarbio (1 mentions)
25 protocols using beyoclick edu 488 kit
1
Quantifying NB Cell Proliferation
2
Assessing Angiogenesis via EdU Labeling
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To evaluate angiogenesis, mice were injected (i.p.) with 5-ethynyl-2′-deoxyuridine (EdU, ST067, Beyotime, Shanghai, China) (50 mg/kg) on days 5, 6, and 7 following injury. EdU incorporation was then assessed using the BeyoClickTM EdU-488 kit (C0071S, Beyotime, Shanghai, China) on 5 µm coronal sections with immunofluorescence.
3
Cell Proliferation Assays for ccRCC
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To perform the CCK-8 test, ccRCC cells were seeded at a density of 2 × 103 cells per well in 96-well plates and cultured in RMPI-1640 medium containing 10% FBS. Based on the manufacturer's instructions, a Cell Counting Kit-8 (Dojindo, Japan) was used to measure the rate of cell growth. ccRCC cells were placed in 24-well plates for the EdU assay. Upon reaching 80% confluence, the BeyoClickTM EdU-488 kit (Beyotime, Shanghai, China) was employed to determine cell growth rate. Following the application of a stain, the cells were observed using a fluorescence microscope manufactured by Olympus in Tokyo, Japan.
4
Quantifying Ovarian Cancer Cell Proliferation
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Ovarian cancer cells (2000 cells/well) were seeded into 96-well plates and received the indicated treatments. Cells were stained with EdU reagent (10 µM) for 2 h at 37 °C. Then, cells were fixed using 4% paraformaldehyde for 15 min, followed by permeabilization with 0.5% Triton X-100 solution for 10 min. Next, the EdU signal was detected using a BeyoClick™ EdU-488 Kit (C0071S, Beyotime) with a fluorescence microscope. DAPI was used to stain nuclei.
5
Proliferation and Migration Assays for ASCs
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ASCs with different treatments were trypsinized and seeded in 96-well culture plates at 6 × 103 cells/well. After ASCs had been cultured overnight, they were subjected to analyses of proliferation using the BeyoClick™EDU-488 kit (C0071S, Beyotime, Shanghai, China). The relative proliferation ratio of treatment group was calculated relative to the NC group. Subsequently, lower chambers were filled with 650 µl Dulbecco’s modified Eagle medium and 3 × 104 ASCs were evenly seeded in the upper chambers of Transwell plates (8.0 µm pore size, Corning, USA). After ASCs had been cultured for 24 h, cells that migrated to the lower surface of the chamber were stained with crustal violet (G1062, Solarbio, Beijing, China) and counted. The relative migration rate represents the ratio of treatment group to the NC group.
6
Cell Viability and Proliferation Assay
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Cell viability was measured by CCK-8 kit (GlpBio, USA). Cells were seeded at 2 × 103 per well into 96-well plates for 24 h and then starved with 1% serum overnight followed by incubation with MIF. The cells were incubated with CCK-8 solution (1:10) for 3 h, and then the absorbance was measured at 450 nm using a microplate reader (Bio‐Rad, USA). The incorporation rate of EdU (5-ethynyl-2' -deoxyuridine) was assayed using the BeyoClick™ EdU-488 Kit (Beyotime, China) according to the manufacturer's instructions. Briefly, EDU was added to the culture medium at a final concentration of 10 μM for 2 h, followed by 15 min of fixation and 10 min of permeabilization at room temperature. After incubation in 0.5 ml Click reaction solution for 30 min protected from light, nuclear staining was performed using Hoechst 33,342 for 10 min. Then the images were performed using an inverted fluorescence microscope and the number of EdU-positive cells/total cells were counted using Image J software (NIH, USA).
7
Measuring Colon Cell Proliferation with BeyoClick™ EdU-488
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BeyoClick™ EdU-488 Kit (Beyotime, Haimen, China) was used to measure the cell proliferation in colon. Briefly, 4 h before rats were sacrificed, they were injected intraperitoneally with 5 mg/kg EdU solution and then the colon was collected and rinsed with saline. Colon samples were then processed using the BeyoClick™ EdU-488 Kit according to the manufacturer’s instructions [21 (link)]. EdU-positive cells were detected with a confocal laser scanning microscope (LSM 710, ZEISS, Germany).
8
EdU Staining for Cellular Proliferation
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EdU staining was performed by using a commercial BeyoClick™ EdU-488 Kit (Beyotime, Shanghai, China). BmN cells were treated with spermidine at 10 μM or inhibitors at 50 μM (DFMO and MGBG mixtures in a molar ratio of 1:1) for 48 h and stained with 10 μM EdU for 2 h at 27 °C. The cells were fixed with stationary liquid for 15 min and then incubated with Click Additive Solution for 30 min behind the scenes. Subsequently, cells were stained with Hoechst 33,342 (Beyotime, Shanghai, China) and immersed in 1× PBS buffer. Finally, fluorescence signals were captured by a fluorescent microscope (Z16, Leica, Wetzlar, Germany). To quantify the EdU-positive cells (green), cells were analyzed by selecting three different fluorescent fields and counted by the counting function of Photoshop CS6 software (Adobe Systems Incorporated, San Jose, CA, USA).
9
Proliferation of Colon Cells
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The proliferation of colon cells was detected by the BeyoClick™ EdU-488 Kit (Beyotime, Haimen, Jiangsu, China). Briefly, 4 h before rats were sacrificed, they were injected intraperitoneally with 5 mg/kg EdU solution and then the colon was collected and rinsed with saline. Colon samples were then processed using the BeyoClick™ EdU-488 Kit according to the manufacturer’s instructions. The EdU positive cells were detected with a confocal laser scanning microscope (LSM 710, ZEISS, Germany).
10
Evaluating Cytotoxic Efficacy of SN38 and RAPA
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Hepa1-6 cells and SNU-449 cells were seeded in 48-well plates at a density of 5000–10,000 cells/well for an overnight incubation. Then, cells were incubated with SN38 (0.05 μg mL−1 and 0.5 μg mL−1 in Hepa1-6 cells and SNU-449 cells, respectively), RAPA (0.1 μg mL−1 and 1 μg mL−1 in Hepa1-6 cells and SNU-449 cells, respectively), SN38&RAPA, SNPs (0.05 μg mL−1 and 0.5 μg mL−1 SN38-equivalent concentrations in Hepa1-6 cells and SNU-449 cells, respectively), and SRNPs (0.05 μg mL−1 SN38-equivalent concentration and 0.1 μg mL−1 RAPA in Hepa1-6 cells and 0.5 μg mL−1 SN38-equivalent concentration and 1 μg mL−1 RAPA in SNU-449 cells) for 48 h. After incubation, cells were further incubated with EdU (5-ethynyl-2′-deoxyuridine) solution (50 μM) (BeyoClick™ EdU 488 Kit, Cat# C0071S, Beyotime, China) for another 2 h. Next, cells were washed and fixed by 4% formaldehyde for 15 min at room temperature. After incubation with 0.3% Triton X-100 for 10 min, cells were treated with Click solution for another 30 min. Hoechst 33,342 was further used to stain the nuclei for 10 min. Finally, the cell proliferation abilities were assessed by fluorescence microscope.
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