Acridine Orange zinc chloride salt  (Synonyms: Euchrysine 3RX)

Acridine Orange (Euchrysine 3RX) zinc chloride salt is a cell-penetrable nucleic acid-selective fluorescent dye. Acridine Orange zinc chloride salt produces orange fluorescence when it binds to ssDNA or RNA, and green fluorescence when it binds to dsDNA (Ex: 488 nM; Em: green fluorescence at 530 nm, orange fluorescence at 640 nm)^[1][2][3].

Guide (Following is our recommended protocol. This protocol only provides a guideline, and should be modified according to your specific needs).
1. Cell Counting with AO/PI (Live/Dead Assay)
(1) Prepare Sample: Mix equal volumes of cell suspension and 1-5 μg/mL Acridine Orange in PBS.
(2) Incubate: No incubation is typically required for simple counting; however, 5-15 minutes in the dark can improve results.
(3) Analyze: Use a fluorescence microscope with blue excitation (e.g., 488 nm).
(4) Result Interpretation: Live nucleated cells display green fluorescence, while dead cells with compromised membranes display red fluorescence.
2. Microbial Staining (Slide Protocol)
(1) Prepare Sample: Prepare a smear on a clean glass slide and allow to air dry.
(2) Fixation: Fix the smear using methanol (2-3 minutes).
(3) Staining: Flood the slide with 0.1% Acridine Orange stain and allow it to sit for 2 minutes.
(4) Rinse & Decolorize: Rinse with tap water, then wash with alcoholic saline (10% ethanol in 0.9% NaCl) for 1-2 minutes to reduce background.
(5) Imaging: Air dry and examine using a fluorescence microscope.
3. Cell Cycle Analysis (Flow Cytometry)
(1) Suspension: Aliquot 10⁵-10⁶ cells in 100 μl PBS.
(2) Permeabilization: Add 0.5 mL of Buffer I (0.1% Triton X-100, 0.2M Sucrose, 0.1M Citric Acid) at room temperature.
(3) Staining: Add 0.5 mL of Staining Buffer II (0.1 M NaCl, 0.1 M Sodium Phosphate/Citrate buffer, pH 3.8, with 20 μg/mL AO).
(4) Analyze: Immediately run on a flow cytometer.

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

Acridine Orange (0.1 mg/kg, i.v., dogs) zinc chloride salt shows no clinical signs of toxicity and no abnormalities were seen in the blood within 30 days^[1].

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

438.10

C₁₇H₂₀Cl₃N₃Zn

10127-02-3

Solid

Brown to reddish brown

Cl[Zn]Cl.CN(C)C1=CC=C2C(N=C3C=C(N(C)C)C=CC3=C2)=C1.Cl

Room temperature in continental US; may vary elsewhere.

4°C, sealed storage, away from moisture and light

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

* Please refer to the solubility information to select the appropriate solvent. Once prepared, please aliquot and store the solution to prevent product inactivation from repeated freeze-thaw cycles.
Storage method and period of stock solution: -80°C, 6 months; -20°C, 1 month (sealed storage, away from moisture and light). When stored at -80°C, please use it within 6 months. When stored at -20°C, please use it within 1 month.

* Note: If you choose water as the stock solution, please dilute it to the working solution, then filter and sterilize it with a 0.22 μm filter before use.

• [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.  [Content Brief]

  [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.  [Content Brief]

  [3]. Byvaltsev VA, et al. Acridine Orange: A Review of Novel Applications for Surgical Cancer Imaging and Therapy. Front Oncol. 2019 Sep 24;9:925.  [Content Brief]

  [4]. Wang Q, et al. Substrate stiffness regulates the differentiation profile and functions of osteoclasts via cytoskeletal arrangement. Cell Prolif. 2022 Jan;55(1):e13172.  [Content Brief]