Propidium iodide (PI) is a red-fluorescent nuclear and chromosome counterstain which can bind to double stranded DNA by intercalating between base pairs.
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描述 | Propidium iodide (PI) is a red-fluorescent nuclear and chromosome counterstain which can bind to double stranded DNA by intercalating between base pairs. It is a cell impermeant dye, which can be excluded from viable cells. Thus it is widely used for the evaluation of apoptosis in different experimental models. Propidium iodide is excited at 488 nm and, with a relatively large Stokes shift, emits at a maximum wavelength of 617 nm. General protocol: 1. Trypsinize and harvest cells in the appropriate manner and wash in PBS. 2. Fix cells into 0.5 ml 70% EtOH for at least 1h. 70% EtOH should be pre-cooled to -20 °C overnight. PAUSE POINT: Cells can be stored in ethanol solution at –20°C for several weeks. 3. Spin down cells for 2 min at 2,000 rpm. 4. Discard the supernatant. Be careful to avoid cell loss. 5. Wash in PBS for two times as: resuspend cell pellet in 0.5mL 1xPBS, spin down cells for 2 min at 2,000 rpm, discard the supernatant and repeat these steps. 6. Resuspend cell pellet in 0.5 ml PBS containing 10 μg/ml RNase A and 20 μg/ml PI stock solution, transfer to FACS tubes. Note: The addition of RNase A could ensure that only DNA is stained. 7. Incubate at room temperature in the dark for 30 min. 8. Analyze cells by flow cytometry. Use 488-nm laser line for excitation. Measure red fluorescence (>600 nm) and side scatter. Optimal protocol: Fluorochrome solution: 0.1% sodium citrate (wt/v), 0.1% Triton X-100 |
计算器 | ||||
存储液制备 | ![]() |
1mg | 5mg | 10mg |
1 mM 5 mM 10 mM |
1.50mL 0.30mL 0.15mL |
7.48mL 1.50mL 0.75mL |
14.96mL 2.99mL 1.50mL |
CAS号 | 25535-16-4 |
分子式 | C27H34I2N4 |
分子量 | 668.395 |
别名 | 碘化丙啶 ;PI |
运输 | 蓝冰 |
存储条件 |
In solvent -20°C:3-6个月-80°C:12个月 Pure form Keep in dark place,Sealed in dry,2-8°C |
溶解方案 |
DMSO: 105 mg/mL(157.09 mM),配合低频超声助溶,注意:DMSO长时间开封后,会吸水并导致溶解能力下降,请避免使用长期开封的DMSO H2O: 3 mg/mL(4.49 mM),配合低频超声,并水浴加热至45℃助溶 以下溶解方案都请先按照体外实验的方式配制澄清的储备液,再依次添加助溶剂: ——为保证实验结果的可靠性,澄清的储备液可以根据储存条件,适当保存;体内实验的工作液,建议现用现配,当天使用; 以下溶剂前显示的百分比是指该溶剂在终溶液中的体积占比;如在配制过程中出现沉淀、析出现象,可以通过加热和/或超声的方式助溶
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动物实验配方 |