Propidium Iodide is a red-fluorescent dye that can be used to stain cells.

Propidium Iodide is a cell-membrane impermeable dye with characteristic excitation maximum at 535 nm and emission maximum at 617 nm which intercalates with nucleic acids with a stoichiometry of one dye per 4-5 base pairs with little sequence preference. Propidium Iodide has evidenced of having no toxic effects on neurons, being today’s most common marker for membrane integrity and cell viability when applied prior to fixation (pre-fixation Propidium Iodide staining method). The pre-fixation staining has been widely used for quantitative assessments of neuronal cell decline in models of acute neurodegeneration, visualized as intensely labeled PI⁺-pycnotic nuclei of degenerating neurons ^[1]. Propidium Iodide cannot cross the membrane of live cells, making it useful to measure the percentage of apoptotic cells by flow-cytometric analysis. The flow cytometric data shows an excellent correlation with the results obtained with both electrophoretic and colorimetric methods. This new rapid, simple and reproducible method proves useful for assessing apoptosis of specific cell populations in heterogeneous tissues such as bone marrow, thymus and lymph nodes^[2].

MCE has not independently confirmed the accuracy of these methods. They are for reference only.

668.39

C₂₇H₃₄I₂N₄

25535-16-4

碘化丙啶

Room temperature in continental US; may vary elsewhere.

4deg;C, sealed storage, away from moisture and light

*In solvent : -80deg;C, 6 months; -20deg;C, 1 month (sealed storage, away from moisture and light)

DMSO : 100 mg/mL (149.61 mM; Need ultrasonic)

H₂O : 3.57 mg/mL (5.34 mM; ultrasonic and warming and heat to 60°C)

请根据产品在不同溶剂中的溶解度选择合适的溶剂配制储备液；一旦配成溶液，请分装保存，避免反复冻融造成的产品失效。
储备液的保存方式和期限：-80°C, 6 months; -20°C, 1 month (sealed storage, away from moisture and light)。-80°C 储存时，请在 6 个月内使用，-20°C 储存时，请在 1 个月内使用。

请根据您的实验动物和给药方式选择适当的溶解方案。以下溶解方案都请先按照 In Vitro 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，建议您现用现配，当天使用； 以下溶剂前显示的百
分比是指该溶剂在您配制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

• 1.

  请依序添加每种溶剂：;10% DMSO ;; 40% PEG300 ;; 5% Tween-80 ;; 45% saline

  Solubility: ≥ 2.5 mg/mL (3.74 mM); Clear solution

  此方案可获得 ≥ 2.5 mg/mL (3.74 mM，饱和度未知) 的澄清溶液。

  以 1 mL 工作液为例，取 100 μL 25.0 mg/mL 的澄清 DMSO 储备液加到 400 μL PEG300 中，混合均匀；向上述体系中加入50 μL Tween-80，混合均匀；然后继续加入 450 μL生理盐水定容至 1 mL。

  将 0.9 g 氯化钠，完全溶解于 100 mL ddH₂O 中，得到澄清透明的生理盐水溶液

• 2.

  请依序添加每种溶剂：;10% DMSO ;; 90% (20% SBE-β-CD in saline)

  Solubility: ≥ 2.5 mg/mL (3.74 mM); Clear solution

  此方案可获得 ≥ 2.5 mg/mL (3.74 mM，饱和度未知) 的澄清溶液。

  以 1 mL 工作液为例，取 100 μL 25.0 mg/mL 的澄清 DMSO 储备液加到 900 μL 20% 的 SBE-β-CD 生理盐水水溶液中，混合均匀。

  将 2 g 磺丁基醚 β-环糊精加入 5 mL 生理盐水中，再用生理盐水定容至 10 mL，完全溶解，澄清透明

• [1]. Hezel M, et al. Propidium iodide staining: a new application in fluorescence microscopy for analysis of cytoarchitecture in adult and developing rodent brain. Micron. 2012 Oct;43(10):1031-8.

  [2]. A rapid and simple method for measuring thymocyte apoptosis by propidium iodidestaining and flow cytometry. J Immunol Methods. 1991 Jun 3;139(2):271-9.

Flow cytometric analysis: Propidium iodide is prepared in in 0.1% sodium citrate plus 0.1% Triton X-100 (50 μg/mL). The 200 ×g centrifuged cell pellet is gently resuspended in 1.5 mL hypotonlc fluorochrome solution (Propidium iodide 50 μg/mL), in 12×75 polypropylene tubes. The tubes are placed at 4°C in the dark overnight before the flow cytometric analysis. The propidium Iodide fluorescence of individual nuclei is measured using a FACScan flow cytometer. The nuclei traverses the light beam of a 488 nm Argon laser. A 560 nm dichrolc nurror (DM 570) and a 600 nm band pass filter (bandwidth 35 nm) are used for collecting the red fluorescence due to propidium Iodide staining of DNA and the data are registered on a logarithmic scale^[2].

MCE has not independently confirmed the accuracy of these methods. They are for reference only.

• [1]. Hezel M, et al. Propidium iodide staining: a new application in fluorescence microscopy for analysis of cytoarchitecture in adult and developing rodent brain. Micron. 2012 Oct;43(10):1031-8.

  [2]. A rapid and simple method for measuring thymocyte apoptosis by propidium iodidestaining and flow cytometry. J Immunol Methods. 1991 Jun 3;139(2):271-9.