Dulbecco’s Modified Eagle’s Medium (DMEM, Hyclone, Illinois, USA), trypsin (Hyclone, USA), phosphate-buffered saline (PBS, Hyclone, USA), fetal bovine serum (FBS, BI, Israel, USA), dimethyl sulfoxide (Sigma, St. Louis, Missouri, USA), a Cell Counting Kit-8 (CCK-8 Kit) (Dojindo, Kumamoto, Japan), cell cycle staining buffer (Multi Sciences, Hangzhou, China), Hoechst 33342/PI double staining kit (Solarbio, Beijing, China), antibodies (Abcam, Massachusetts, USA), petri dishes and culture flasks (Corning, New York, USA), and conventional reagents of analytical pure grade (Sinopharm, Shanghai, China).
Hoechst 33342 pi double staining kit
Manufactured by Solarbio
Sourced in China
The Hoechst 33342/PI double staining kit is a laboratory tool used for simultaneous staining and visualization of cellular DNA. The kit contains Hoechst 33342, a blue-fluorescent dye that binds to DNA, and propidium iodide (PI), a red-fluorescent dye that binds to DNA. This combination allows for the differentiation of live and dead cells in a sample.
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5 protocols using hoechst 33342 pi double staining kit
1
Cell Proliferation and Cell Cycle Analysis
2
Apoptosis Detection via Hoechst-PI Staining
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On the basis of the relationship between cell apoptosis and membrane permeability, Hoechst 33342-PI double staining kit (solarbio, Beijing, China) was utilized to detect cell apoptosis. The SCC-15 and Cal-27 cells received designed treatment, followed by staining via suitable amount of Hoechst 33342-PI mixed dye fluid for 30 minutes in dark. Eventually, a fluorescence microscope (Leica Dmi8, Wetzlar, Germany) was an effective method to record representative images.
3
Hoechst 33,342 and PI Dual Staining
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PI/Hoechst 33,342 staining was performed using a Hoechst 33,342/PI Double Staining Kit (Solarbio Science, Beijing China). The cells were evenly grown on the bottom of the culture dish, fixed with 4% paraformaldehyde at 37°C for 15 min, and stained with the dye solution at room temperature and away from light for 30 min. Finally, images of the staining were captured by a confocal laser scanning microscope (FV10i, Olympus, Tokyo, Japan).
4
Quantifying Cell Survival and Death
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A Hoechst 33342/PI double-staining kit (Solarbio, Beijing, China) was used to evaluate cell survival, necrosis, and apoptosis according to the manufacturer’s instructions. HepG2 and HuH7 cells were seeded into six-well plates (5 × 105 cells/well) and cultivated at 37 °C with 5% CO2 overnight. After washing twice with PBS, the cells were stained with Hoechst 33342 for 10 min and PI for another 10 min. After 30 min of incubation, the cells were washed with PBS, and images were captured under a microscope (CKX53; Olympus, Tokyo, Japan).
5
Hoechst 33342/PI Nuclear Staining
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PI/Hoechst 33342 staining was performed using Hoechst 33342/PI Double Staining Kit (Solarbio Science, Beijing China). Cells were plated into a glass bottom cell culture dish at the density of 2.0 × 105. Briefly, the cells were fixed with 4% paraformaldehyde for 15 minutes at 37℃. The stain solution was added into cell culture medium for 30 minutes at room temperature in the dark. Finally, images of the staining were captured by the confocal laser scanning microscope (FV10i, Olympus, Tokyo, Japan).
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