An EdU assay was conducted utilizing an EdU immunofluorescence staining kit (Ribobio). The sterilized slides were put into a 12‐well plate, and then around 1 × 103 cells were seeded into each well. The transfected cells were cultured with 50 μmol/L EdU reagent for 2 hours and washed with PBS (HyClone), followed by fixation with 4% phosphate‐buffered paraformaldehyde. Afterwards the cells were stained with 100 μL of fresh Apollo reaction cocktail, and the nucleus was stained with 100 μL of Hoechst 33 342, followed by viability determination with a fluorescence microscope (Olympus BX51).
Edu immunofluorescence staining kit
Manufactured by RiboBio
Sourced in China
The EdU immunofluorescence staining kit is a product designed to detect and visualize DNA synthesis and cell proliferation. It utilizes the incorporation of the thymidine analog EdU (5-Ethynyl-2'-deoxyuridine) into newly synthesized DNA, which can then be detected using a fluorescent dye. This kit provides a simple and efficient method for labeling and identifying proliferating cells.
Automatically generated - may contain errors
6 protocols using edu immunofluorescence staining kit
1
EdU Immunofluorescence Staining Protocol
2
EdU Cell Proliferation Assay
Check if the same lab product or an alternative is used in the 5 most similar protocols
+ Expand
EdU detection was performed by using an EdU immunofluorescence staining kit (Ribobio, Guangzhou, China) according to the manufacturer’s protocol. Sterilized slides were put into 24 well plates, and H9C2 cells were seeded into each well at a density of 3 × 104 cells/well. Transfected cells were incubated with 20 μmol/L EdU reagent for 2 h, washed twice with phosphate Buffer solution (PBS) and fixed with 4% phosphate buffered paraformaldehyde for 15 min. Subsequently, the cells were stained with 100 μL of fresh Apollo reaction cocktail, and nuclei were stained with 100 μL of Hoechst 33342. The percentage of EdU positive cells was counted by using fluorescence microscope (LEICA).
3
EdU Proliferation Assay Protocol
Check if the same lab product or an alternative is used in the 5 most similar protocols
+ Expand
Transfected cells were seeded into 96-well plates at a density of 1 × 105 cells/mL and cultured overnight with 5% CO2 at 37 °C. EdU assay was performed by the EdU immunofluorescence staining kit (Ribobio). In short, cells of each group were first incubated with 100 μL of EdU reagent for 4 h. Following incubation, the culture medium was discarded, and cells were washed twice with PBS. Next, cells were fixed with 4% paraformaldehyde for 15 min and stained with 5 μg/mL Hoechst 33342 for 30 min. Stained cells were visualized and imaged under a fluorescence microscope (Olympus BX 60 fluorescence microscope).
4
EdU Staining Assay for Fenofibrate Effects
Check if the same lab product or an alternative is used in the 5 most similar protocols
+ Expand
5-Ethynyl-2′-deoxyuridine (EdU) staining assay was carried out on fenofibrate-treated cells utilizing an EdU immunofluorescence staining kit (Ribobio, China) according to the manufacturer's instructions [29 (link)]. The results were observed using an inverted fluorescence microscope (200x).
5
Cell Proliferation Assay: CCK-8 and EdU
Check if the same lab product or an alternative is used in the 5 most similar protocols
+ Expand
The Cell Counting Kit-8 (CCK-8, Dojindo, Kyushu, Kumamoto, Japan) assay was used to examine cell proliferation according to the manufacturer's instructions. After 0 h, 24 h, 48 h, 72 h, and 96 h, CCK-8 was added to the wells and incubated for 2 h at 37°C. Next, the absorbance at 450 nm was detected with an automatic micro-plate reader (Bio-Rad, Hercules, CA, USA) with the purpose of plotting the growth curve. To visually observe the proliferative cells, an EdU incorporation experiment was conducted using an EdU immunofluorescence staining kit (Ribobio, Guangzhou, China). The cells were seeded into 96-well culture plates at a density of 4 × 103 cells/well. EdU medium (1:1,000) was add to the wells and incubated at 37°C for 2 hours. Cells were cleaned by phosphate buffer solution (PBS; Hyclone, Logan, Utah, USA) before being fixed in 4% phosphate-buffered paraformaldehyde. Then cells were stained with fresh Apollo solution and the nucleus was stained with Hoechst33342. Finally, cells were observed with a microscope (Olympus BX51).
6
Proliferative Cell Analysis by EdU and CCK-8
Check if the same lab product or an alternative is used in the 5 most similar protocols
+ Expand
To visually observe the proliferative cells, an EdU (5-ethynyl-2′-deoxyuridine) incorporation experiment was performed using an EdU immunofluorescence staining kit (RiboBio). The cells were collected and seeded into a 96-well culture plate at a density of 4 × 10 3 cells/well. Three repeated wells were set in every group. Then, a transfection assay was performed according to procedures described above. EdU medium (1:1000) was added to wells and incubated at 37°C for 2 h. Cells were rinsed with phosphate buffer solution (PBS) (Hyclone) before being fixed in 4% phosphate-buffered paraformaldehyde. Then, cells were stained with fresh Apollo solution, and nuclei were stained with Hoechst33342. Finally, cells were observed with a microscope (Olympus BX51). Meanwhile, a Cell Counting Kit-8 (CCK-8, Dojindo, Kyushu, Kumamoto, Japan) assay was used to examine cell proliferation according to the manufacturer's instructions. After 0, 24, 48, 72, and 96 h, CCK-8 was added to the wells and incubated for 1 h at 37°C. Next, the absorbance at 450 nm was detected with an automatic microplate reader (Bio-Rad) for the purpose of plotting the growth curve.
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!