Cell proliferation was measured using an EdU staining assay. The EdU detection kit was purchased from Guangzhou RiboBio (cat. no. C10310-1). Firstly, HLECs were cultured in CM or CM-TNF-α for 48 h. Next, HLECs were incubated with 100 μl of 50 μm EdU for 1 h, washed with PBS, and incubated with 1 mg/ml DAPI for 10 min. Next, the EdU-positive HLECs were measured using a fluorescence microscope (IX51; Olympus) [31 (link)].
Edu detection kit
Manufactured by RiboBio
Sourced in China, Japan
The EdU detection kit is a laboratory tool used to identify and quantify cell proliferation. It employs the incorporation of the nucleoside analog 5-ethynyl-2'-deoxyuridine (EdU) into actively dividing cells, which can then be detected using a copper-catalyzed click reaction. The kit provides the necessary reagents and protocols to perform this analysis.
Automatically generated - may contain errors
Lab products found in correlation
Fluorescence microscope, by Olympus (5 mentions)
Hoechst 33342 dye, by Thermo Fisher Scientific (3 mentions)
Triton x 100, by Merck Group (2 mentions)
Cell cycle detection kit, by BestBio (2 mentions)
Fetal bovine serum (fbs), by Thermo Fisher Scientific (2 mentions)
P akt, by Abcam (2 mentions)
Annexin 5 pi kit, by BestBio (2 mentions)
Emodin, by Yuanye Bio-Technology (2 mentions)
Goat anti rabbit igg, by Proteintech (2 mentions)
Ad mcherry gfp lc3, by RiboBio (2 mentions)
Chloroquine cq, by Selleck Chemicals (2 mentions)
P pi3k, by Abcam (2 mentions)
Trizol, by Thermo Fisher Scientific (2 mentions)
Dimethyl sulfoxide (dmso), by Merck Group (2 mentions)
Vimentin, by Abcam (2 mentions)
Matrigel, by Corning (2 mentions)
E cadherin, by Abcam (2 mentions)
N cadherin, by Abcam (2 mentions)
Fluorescence microscope, by Leica (2 mentions)
Microplate reader, by Bio-Rad (2 mentions)
63 protocols using edu detection kit
1
Quantifying HLEC Proliferation via EdU Assay
2
EdU-Based Cell Proliferation Assay
Check if the same lab product or an alternative is used in the 5 most similar protocols
+ Expand
The EdU incorporation assay was carried out with an EdU detection Kit (RiboBio, Guangzhou, China) to assess cell proliferation viability according to the manufacturer’s protocol. All images were acquired with an Olympus IX73-FL-PH fluorescence microscope (Olympus, Tokyo, Japan). The experiments were performed at least three times independently with triplicate samples.
3
Glioma Cell Colony Formation and Proliferation Assays
Check if the same lab product or an alternative is used in the 5 most similar protocols
+ Expand
For colony formation assay, the glioma cells of U251 and A172 were harvested and cultured in six-well plates at a thickness of 300 cells/well at 37°C for incubation. After culturing for 14 days, the forming colony cells were fixed with paraformaldehyde (4%, Beyotime, Beijing, China) for 10 min, stained with crystal violet (0.5%, Beyotime) for 30 min, and washed three times with phosphate-buffered saline (PBS). The number of visible colonies was photographed, manually recorded, and statistically analyzed.
Following transfection, U251 and A172 cells were cultured in 96-well plates at a thickness of 3 × 104 cells/well and incubated for 48 h. Next, 50 µmol/L 5-ethynyl-2′-deoxyuridine (EdU) labeling solution was added to the culture of each well for 2 h. Then, the cells were fixed with a 4% fixative solution (Sigma-Aldrich) for 30 min and washed twice with PBS. Afterward, 0.5% TritonX-100 (Sigma-Aldrich) was added and stained with the EdU detection kit (RiboBio, Guangzhou, China). Finally, the green-colored positive cells and blue-colored nuclei were observed and photographed using a fluorescence microscope (Olympus, Tokyo, Japan). The proliferative ability of cells was determined by calculating the positive cell rates.
Following transfection, U251 and A172 cells were cultured in 96-well plates at a thickness of 3 × 104 cells/well and incubated for 48 h. Next, 50 µmol/L 5-ethynyl-2′-deoxyuridine (EdU) labeling solution was added to the culture of each well for 2 h. Then, the cells were fixed with a 4% fixative solution (Sigma-Aldrich) for 30 min and washed twice with PBS. Afterward, 0.5% TritonX-100 (Sigma-Aldrich) was added and stained with the EdU detection kit (RiboBio, Guangzhou, China). Finally, the green-colored positive cells and blue-colored nuclei were observed and photographed using a fluorescence microscope (Olympus, Tokyo, Japan). The proliferative ability of cells was determined by calculating the positive cell rates.
4
Cell Cycle and Proliferation Analysis
Check if the same lab product or an alternative is used in the 5 most similar protocols
+ Expand
FACS and flow cytometry analysis were performed according to the standard protocol. For cell cycle analysis, each 1 ml of cell suspension (1–5 × 105 cells) was incubated with the (propidium iodide (PI), Cell Signalling Technology). For EdU analysis, 1–5 × 105 cells from each sample were processed with EdU (EdU Detection Kit, Ribobio), in accordance with the manufacturer's instructions.
5
EdU Proliferation Assay Protocol
Check if the same lab product or an alternative is used in the 5 most similar protocols
+ Expand
An EdU detection kit (C10310, Guangzhou RiboBio Co., Ltd., Guangdong, China) was employed for the proliferation assay. The 1 × 104 cells were seeded in 96-well plate and incubated for 24 h. Then, 100 µl fresh medium containing 50 µM EdU was added into the plate and incubated at 37 °C for 2 h. After this incubation, cells were fixed by addition of 20 g/l PFA for 20 min, de-crosslinked twice by addition of 2 mg/ml glycin, and incubated with PBST for 10 min, followed by incubation with 100 µl Apollo staining solution for 30 min. After that, cells were washed twice with PBS, treated with Hoechst33342 for 30 min in the dark, and washed in 0.5% Triton X-100. Finally, the cells were observed under a fluorescence microscope and counted with the Image-pro plus 6.0 software.
6
Evaluating cell proliferation with EdU
Check if the same lab product or an alternative is used in the 5 most similar protocols
+ Expand
The EdU assay was carried out with the EdU detection kit (RiboBio, Guangzhou, China). 2 × 103 PASMCs were seeded into 96-well plates in the logarithmic growth phase. Each well was incubated with 50 µM EdU solution for 2 h and washed with PBS twice for 5 min. 4% paraformaldehyde was used to fix PASMCs at room temperature for 30 min, and the cells were incubated in 1 × Apollo staining solution for 30 min at room temperature. Finally, 1 × DAPI solution was added to each well and incubated at room temperature in the dark for 30 min. Fluorescence was detected using a fluorescence microscope (Olympus Corporation, Japan).
7
EdU-based Cell Proliferation Assay
Check if the same lab product or an alternative is used in the 5 most similar protocols
+ Expand
Cell proliferation analysis was carried out with an EdU detection kit (RiboBio, Guangzhou, China). Cells were cultured with 50 μM EdU for 2 h at 37°C and then fixed in 4% paraformaldehyde for 30 min and permeabilized with 0.5% Triton X-100 for 10 min. Afterwards, cells were treated with Apollo® reaction cocktail for 30 min. For nuclear staining, cells were treated with Hoechst 33342 for 30 min and visualized with a fluorescent microscope (Olympus, Tokyo, Japan).
8
Quantifying Cell Proliferation Using EdU Assay
Check if the same lab product or an alternative is used in the 5 most similar protocols
+ Expand
An EdU detection kit (RiboBio) was used to detect cell proliferation. H460 and A549 cells were cultured for 24 h. The cells were then treated with 50 μM EdU at 37°C for 2 h. Next, the culture solution was discarded, and subsequently, the A549 and H460 cells were fixed for 30 min with 4% paraformaldehyde. Next, 0.5% Triton X-100 was applied to increase the permeability, and then, the A549 and H460 cells were incubated with Apollo fluorescence staining reaction solution for 30 min in a dark place. Then, the cells were stained for 15 min with DAPI staining solution. Finally, the cells were placed under a fluorescence microscope (magnification of 200) (Olympus), and the EdU-positive cells were counted.
9
Senescence-Associated Beta-Galactosidase Assay and EdU Proliferation Analysis
Check if the same lab product or an alternative is used in the 5 most similar protocols
+ Expand
SA-β-gal staining was performed to determine the level of SA-β-gal expression in U2OS cells using a kit purchased from Sigma‒Aldrich (Germany) according to the manufacturer's instructions. Briefly, U2OS cells were harvested and fixed with fixation buffer, washed three times in PBS and permeabilized prior to staining. After that, the cells were stained with SA-β-gal staining solution overnight at 37 °C. Finally, SA‐β‐gal‐positive cells, stained blue, were randomly imaged. For cell proliferation analysis, EdU detection kit (RiboBio, C10310) was used to measure the difference in live cell count and cell percentages between different groups. An overnight incubation with EdU reagent solution at 37 °C followed by 10 min of fixation with 4% paraformaldehyde was carried out in U2OS cells. The following day, the cells were subjected to imaging analysis and cell counting. For statistical analysis, this formula calculated the percentage of EdU-positive cells based on the count of EdU-positive cells (EdU-positive cells + EdUu-negative cells).
10
Quantifying Cell Proliferation with EdU
Check if the same lab product or an alternative is used in the 5 most similar protocols
+ Expand
Cultured cells were stained with EdU detection kit (Ribobio, Guangzhou, China) following the manufacturer’s instruction. Briefly, cells were incubated with 50 μM EdU labeling medium at 37°C for 2h in 5% CO2 incubator. After immobilization using 500 μL 4% paraformaldehyde at room temperature for 10 min, cells were stained with Apollo®567 solution and Hoechst33342 solution. Cells were observed under a X71 (U-RFL-T) fluorescence microscope (Olympus, Melville, NY). EdU-positive cells represent proliferating cells and Hoechst33342-positive cells represent total cells.
About PubCompare
Our mission is to provide scientists with the largest repository of trustworthy protocols and intelligent analytical tools, thereby offering them extensive information to design robust protocols aimed at minimizing the risk of failures.
We believe that the most crucial aspect is to grant scientists access to a wide range of reliable sources and new useful tools that surpass human capabilities.
However, we trust in allowing scientists to determine how to construct their own protocols based on this information, as they are the experts in their field.
Ready to get started?
Sign up for free.
Registration takes 20 seconds.
Available from any computer
No download required
Revolutionizing how scientists
search and build protocols!