Acridine Orange hydrochloride is a cell-permeable fluorescent dye that binds to nucleic acids, resulting in an altered spectral emission.

Acridine Orange has been employed extensively as a cytochemical stain and has shown to stain differentially DNA and RNA, and double-restranded nucleic acids in situ. Acridine Orange can either intercalate into double helical nucleic acids (green fluorescence at 530 nm), or bind electrostatically to phosphate groups of single-stranded molecules (red fluorescence at 640 nm). This unique characteristic makes acridine orange useful for cell-cycle studies^[1]. Acridine Orange staining of unfixed cells may be used as a simple, fast means of obtaining information on cell ploidy levels and cell cycle status from DNA measurements (green fluorescence), and cell transcriptional activity from RNA staining (red fluorescence), in human and murine cells lines, peripheral blood and bone marrow specimens from patients with leukemia and mitogenically (phytohemagglutinin) or antigenically (mixed lymphocyte culture) stimulated human peripheral blood cultures^[2].

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

301.81

C₁₇H₂₀ClN₃

65-61-2

吖啶橙；碱性橙 14；丫啶橙；3,6-双(二甲氨基)吖啶

Room temperature in continental US; may vary elsewhere.

4deg;C, protect from light, stored under nitrogen

*In solvent : -80deg;C, 6 months; -20deg;C, 1 month (protect from light, stored under nitrogen)

H₂O : 50 mg/mL (165.67 mM; ultrasonic and heat to 60°C)

DMSO : 25 mg/mL (82.83 mM; Need ultrasonic)

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 (protect from light, stored under nitrogen)。-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: 2.08 mg/mL (6.89 mM); Suspended solution; Need ultrasonic

  此方案可获得 2.08 mg/mL (6.89 mM) 的均匀悬浊液，悬浊液可用于口服和腹腔注射。

  以 1 mL 工作液为例，取 100 μL 20.8 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: ≥ 2.08 mg/mL (6.89 mM); Clear solution

  此方案可获得 ≥ 2.08 mg/mL (6.89 mM，饱和度未知) 的澄清溶液。

  以 1 mL 工作液为例，取 100 μL 20.8 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]. McMaster GK, et al. Analysis of single- and double-stranded nucleic acids on polyacrylamide and agarosegels by using glyoxal and acridine orange. Proc Natl Acad Sci U S A. 1977 Nov;74(11):4835-8.

  [2]. Traganos F, et al. Simultaneous staining of ribonucleic and deoxyribonucleic acids in unfixed cells using acridine orange in a flow cytofluorometric system. J Histochem Cytochem. 1977 Jan;25(1):46-56.

Two-step, pH 3.0: Aliquots (0.2 mL, containing approximately 2-5×10⁵ cells) are withdrawn from cultures and are added to 0.5 mL of a solution containing: 0.1% (v/v) Triton X-100, 0.2 M sucrose, 10^-4 M EDTA and 2×10^-2 M citrate-phosphate buffer, at pH 3.0. Triton X-100 is included in the various procedures at the indicated pH to increase cell permeability yet maintain cellular integrity. The chelating agent EDTA is used to facilitate RNA denaturation. The cells are stained one minute later by addition of 1 mL of a solution containing 0.002% (20 μg/mL) AO, 0.1 M NaCl and 10^-2 M citrate-phosphate buffer, pH 3.8. Cations are included in the staining mixture to ensure staining specificity. The final AO concentration is approximately 4×10^-5 M^[2].

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

• [1]. McMaster GK, et al. Analysis of single- and double-stranded nucleic acids on polyacrylamide and agarosegels by using glyoxal and acridine orange. Proc Natl Acad Sci U S A. 1977 Nov;74(11):4835-8.

  [2]. Traganos F, et al. Simultaneous staining of ribonucleic and deoxyribonucleic acids in unfixed cells using acridine orange in a flow cytofluorometric system. J Histochem Cytochem. 1977 Jan;25(1):46-56.