EdU assay was used to examine cell proliferation. Glioma cell lines in the logarithmic growth phase were seeded into 96-well plates at a density of 2×10³- 4×10⁴. After 24 h, the adherent cells to the wells were transfected. Five parallel wells were set up for each group. Cells in each well after transfection for 48 h were cultured with 100 μL EdU medium (RIBOBIO, China) for 2 h and fixed with 100 μL of cell fixation solution (PBS containing 4% polyformaldehyde) for 30 min at room temperature. Subsequently, the cells were incubated with 2 mg/mL glycine (Solarbio, China) for 5 min, rinsed with 100 μL of PBS containing 0.5% TritonX-100 (RIBOBIO, China) for 10 min, and stained using 1 × Apollo staining reaction solution (RIBOBIO, China) for 30 min in conditions devoid of light. Next, the cells were reacted with 100 μL of the 1 × Hoechst 33342 reaction solution (RIBOBIO, China) for 30 min and sealed with 100 μL of the anti-fluorescence quenching agent. Six to ten fields of view were randomly selected for each well and photographed under a fluorescence microscope.
Hoechst 33342 reaction solution
Manufactured by RiboBio
Sourced in China
Hoechst 33342 reaction solution is a fluorescent dye that binds to the minor groove of DNA. It is commonly used in cellular and molecular biology applications for staining nuclei and visualizing DNA in live or fixed cells.
Automatically generated - may contain errors
Lab products found in correlation
Apollo staining reaction solution, by RiboBio (4 mentions)
Edu medium, by RiboBio (3 mentions)
Tritonx 100, by RiboBio (3 mentions)
Glycine, by Solarbio (2 mentions)
Inverted phase contrast microscope, by Olympus (1 mentions)
Glycine, by neoFroxx (1 mentions)
Apollo staining solution, by RiboBio (1 mentions)
Triton x 100, by Beyotime (1 mentions)
5 protocols using hoechst 33342 reaction solution
1
Quantifying Glioma Cell Proliferation
2
Quantifying Cellular Proliferation via EdU Assay
Check if the same lab product or an alternative is used in the 5 most similar protocols
+ Expand
EdU assay was used to examine cell proliferation. Glioma cell lines in the logarithmic growth phase were seeded into 96-well plates at a density of 2×10³-4×10⁴. After 24 h, the adherent cells to the wells were transfected. Five parallel wells were set up for each group. Cells in each well after transfection for 48 h were cultured with 100 μL EdU medium (RIBOBIO, China) for 2 h and xed with 100 μL of cell xation solution (PBS containing 4% polyformaldehyde) for 30 min at room temperature. Subsequently, the cells were incubated with 2 mg/mL glycine (Solarbio, China) for 5 min, rinsed with 100 μL of PBS containing 0.5% TritonX-100 (RIBOBIO, China) for 10 min, and stained using 1 × Apollo staining reaction solution (RIBOBIO, China) for 30 min in conditions devoid of light. Next, the cells were reacted with 100 μL of the 1 × Hoechst 33342 reaction solution (RIBOBIO, China) for 30 min and sealed with 100 μL of the antiuorescence quenching agent. Six to ten elds of view were randomly selected for each well and photographed under a uorescence microscope [9] .
3
Proliferation Assay for CD24(+) and CD24(-) SCAPs
Check if the same lab product or an alternative is used in the 5 most similar protocols
+ Expand
The fifth generation CD24 (+)-SCAPs and CD24 (−)-SCAPs were prepared with good growth condition. After digested with 0.25% trypsin, 100 ul cells (4 × 104/mL) were incubated in a 96-well palatefor 4 h at 37 °C in 5% CO2. Each group of cells were reconstituted in 3 wells. After discarded the culture medium, 150 μL of EdU medium (containing 25 μmol EdU, 10% FBS α-MEM, Hyclone) was added to each well of the cells. After incubating for 48 h at 37 °C in 5% CO2, the cells were washed twice with PBS for 5 min, and 50 μL of 4% paraformaldehyde was added to each well and the cells were fixed for 30 min at room temperature. Then, added 50 μL 2 mg/ml glycine to the cells and shaken for 5 min, and, added 100 μL of penetrant (0.5% Triton X-100) to each well and shaken for 10 min. After washed the cells with PBS for 5 min, 100 μL Apollo staining reaction solution (Ribobio, Guangzhou, China) were added to each well and shaken for 30 min in the dark. After washed 3 times with formaldehyde solution, 100 μL Hoechst 33,342 reaction solution (Ribobio) was added to each well and incubated for 30 min at room temperature. Then washed the cells 5 times, and added 100 μL PBS to each well for observing under an inverted phase-contrast microscope (Olympus).
4
Quantifying Cell Proliferation Using EdU Assay
Check if the same lab product or an alternative is used in the 5 most similar protocols
+ Expand
Cell proliferation assays were performed using the Cell-LightTM EdU Apollo In Vitro Kit (RiboBio, Guangzhou, China). Cancer cells (1 × 105) were seeded in 96-well plates and treated according to the experimental requirements. Subsequently, EdU medium was added and the cells were incubated for 2 h. After washing the cells twice with PBS, they were fixed with 4% PFA solution (Biosharp) for 30 min. Cells were then incubated with glycine (2 mg/ml; Biofroxx, Germany) for 5 min and permeabilized with 0.5% Triton X-100 (Beyotime) for 10 min. After adding 1 × Apollo® staining solution (RiboBio) and incubating for 30 min at light avoidance and RT, the cells were washed three times with 0.5% TritonX-100. Finally, a 1 × Hoechst 33342 reaction solution (RiboBio) was added and incubated for 30 min with light avoidance at RT. The cells were imaged using a microscope (Leica Microsystems) and counted in four randomly selected fields.
5
Quantitative Cell Proliferation Assay with EdU
Check if the same lab product or an alternative is used in the 5 most similar protocols
+ Expand
An EdU assay was used to detect cell proliferation. A total of 2×105 THCA cells/well (HTH-83, K-1, or TPC1) were plated in 96-well plates. After 24 h, the adherent cells were transfected. After 48 h, 100 µl EdU medium containing 10 µM (Guangzhou RiboBio Co., Ltd.) was added to each well for 2 h, and the culture medium was washed and fixed with 100 µl cell fixator (cat. no. P1110; Beijing Solarbio Science & Technology Co., Ltd.) at room temperature for 30 min. The fixed cells were permeabilized with 100 µl PBS containing 0.5% Triton X-100, followed by staining using an Apollo staining reaction solution (Guangzhou RiboBio Co., Ltd.) in the dark for 30 min at 37°C. A Hoechst 33342 reaction solution (100 µl 1×; Guangzhou RiboBio Co., Ltd.) was used for 10 min at 37°C. The dyed plate was placed under an inverted fluorescence microscope (×100 magnification) to obtain fluorescence images.
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!