Cells were collected by trypsinization and washed twice with cold PBS. After fixation in ice-cold 75% ethanol for one hour at -20℃ or overnight at 4℃, cells were washed with cold PBS and resuspended with 500 µL cold PBS. Then 20 µL RNase A solution was added and incubated in a 37 ℃ water bath for 30 minutes. Each sample was stained with 400 µL PI stain solution (BestBio, ShangHai, China) at 4℃ for 60 min in the dark. Cell cycle distributions were detected by Accuri C6 flow cytometer (Becton Dickinson, Franklin Lakes, New Jersey, USA).
Pi stain solution
Manufactured by BestBio
Sourced in China
PI stain solution is a laboratory reagent used for DNA staining and analysis. It binds to DNA and emits fluorescence upon excitation, allowing for the detection and quantification of DNA content in samples.
Automatically generated - may contain errors
Lab products found in correlation
2 protocols using pi stain solution
1
Cell Cycle Analysis by Flow Cytometry
2
Cell Cycle Analysis and Apoptosis Assay
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Cells were collected with 0.25% trypsin and washed twice with cold PBS, then xed with 70%-75% frozen ethanol at -20 °C for 1 h or xed overnight at 4 °C. Subsequently, cells were washed once with cold PBS and then resuspended in 200-500 µl cold PBS. Then, 20 µl RNase A solution was added, and samples were incubated in a 37 °C water bath for 30 min. Finally, 400 µl PI stain solution (BestBio, Shanghai, China) was mixed gently and incubated for 30-60 at 4 °C in the dark. The results were detected with an Accuri C6 ow cytometer (Becton Dickinson, Franklin Lakes, New Jersey, USA).
Apoptosis analyses NCI-H446 and H1299 cells were plated on six-well plates at a concentration of 1 × 10 5 cells/well.
Transfection experiments were performed the following day. Cells were harvested after 48 h and washed twice with PBS. Cells were resuspended in 400 µl 1× buffer, then stained with 5 µl Annexin-V-FITC and 5 µl PI (50 μg/ml) with an Annexin V FITC Apop Dtec Kit (BD Biosciences, San Jose, CA, USA) in the dark for 15 min at room temperature, and detected with an Accuri C6 ow cytometer (Becton Dickinson, Franklin Lakes, New Jersey, USA).
Apoptosis analyses NCI-H446 and H1299 cells were plated on six-well plates at a concentration of 1 × 10 5 cells/well.
Transfection experiments were performed the following day. Cells were harvested after 48 h and washed twice with PBS. Cells were resuspended in 400 µl 1× buffer, then stained with 5 µl Annexin-V-FITC and 5 µl PI (50 μg/ml) with an Annexin V FITC Apop Dtec Kit (BD Biosciences, San Jose, CA, USA) in the dark for 15 min at room temperature, and detected with an Accuri C6 ow cytometer (Becton Dickinson, Franklin Lakes, New Jersey, USA).
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