Acridine Orange hydrochloride

Name: Acridine Orange hydrochloride
Brand: MedChemExpress
SKU: HY-101879
Rating: 5.0 (38 reviews)

Cat. No.: HY-101879 Purity: 99.65%: Data Sheet
COA

SDS
Handling Instructions
FAQs
Technical Support

Acridine Orange hydrochloride is a cell-penetrable nucleic acid-selective fluorescent dye. Acridine Orange hydrochloride 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).

For research use only. We do not sell to patients.

CAS No. : 65-61-2

Size	Stock	Quantity

Free Sample (0.1 - 0.2 mg)	Apply Now
Solid + Solvent (Highly Recommended)
10 mM * 1 mL in DMSO ready for reconstitution	In-stock	Estimated Time of Arrival: December 31
Solution
10 mM * 1 mL in DMSO	In-stock	Estimated Time of Arrival: December 31
Solid
100 mg	In-stock	Estimated Time of Arrival: December 31
200 mg	In-stock	Estimated Time of Arrival: December 31
500 mg	In-stock	Estimated Time of Arrival: December 31
1 g	Get quote
5 g	Get quote

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This product is a controlled substance and not for sale in your territory.

Customer Review

Based on 38 publication(s) in Google Scholar

Other Forms of Acridine Orange hydrochloride:

Acridine Orange hydrochloride (solution) In-stock
Acridine Orange zinc chloride salt In-stock
Acridine Orange base In-stock
Acridine Orange hydrochloride (Standard) Get quote

38 Publications Citing Use of MCE Acridine Orange hydrochloride

Cell Imaging/Staining

: Acridine Orange hydrochloride purchased from MedChemExpress. Usage Cited in: Cell Mol Immunol. 2025 Mar;22(3):282-299. [Abstract]; Human PBMCs (H-PBMCs) and Jurkat cells were treated with OA (5, 50, or 500 μM) or 200 ng/mL PMA for 24 hours, after which the cells were stained with acridine orange (AO).

: Acridine Orange hydrochloride purchased from MedChemExpress. Usage Cited in: Nat Commun. 2023 Jun 30;14(1):3877. [Abstract]; Acridine Orange (AO, 5 μM) was added immediately after the ultrasound treatment and was incubated for 30 min at 37°C before confocal microscopy.

: Acridine Orange hydrochloride purchased from MedChemExpress. Usage Cited in: Acta Pharm Sin B. 2021 Oct;11(10):3178-3192.; The Jurkat cells were treated with LW-218 (8 μmol/L) for 0, 1, 2, and 4 h. Before collecting, the cells were stained with 5 μmol/L Acridine Orange hydrochloride solution for 15 min.

: Acridine Orange hydrochloride purchased from MedChemExpress. Usage Cited in: Cell Death Dis. 2021 Jan 13;12(1):80. [Abstract]; The Jurkat cells treated with 25 μM FV-429 for 12 h were stained with Acridine Orange hydrochloride (5 μM) before collection, and the AO fluorescence was determined by a laser confocal microscope.

: Acridine Orange hydrochloride purchased from MedChemExpress. Usage Cited in: Cell Prolif. 2022 Jan;55(1):e13172. [Abstract]; CLSM images of acridine orange (AO)-stained osteoclasts cultured on PDMS substrates were taken; the cells were stained with 1 μM AO at 37°C for 20 min.

: Acridine Orange hydrochloride purchased from MedChemExpress. Usage Cited in: Cell Death Dis. 2020 Jul 30;11(7):607. [Abstract]; Depending on the acidity, Acridine Orange hydrochloride (AO) causes autophagic lysosomes to appear as orange/red fluorescent vesicles, while it caused nuclei to appear green. AO staining demonstrates that LA dose-dependently increased the numbers of intracellular acidic compartments.

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Biological Activity
Protocol
Purity & Documentation
References
Customer Review

Description

Cellular Effect

Cell Line	Type	Value	Description	References
R3230AC	IC₅₀	2.29 μM Compound: AO.HCl	Phototoxicity in rat R3230AC cells irradiated with halogen lamp pretreated 24 hrs before irradiation by MTT assay Phototoxicity in rat R3230AC cells irradiated with halogen lamp pretreated 24 hrs before irradiation by MTT assay	[PMID: 19692249]

In Vitro

Guide (Following is our recommended protocol. This protocol only provides a guideline, and should be modified according to your specific needs).
1. Working Solution: Dilute Acridine Orange hydrochloride stock in serum-free medium or PBS to a final concentration of 1-10 μM or 2-5 μg/mL.
2. Incubation: Incubate cells for 15-30 min at 37 ℃.
3. Washing: Wash cells twice with pre-warmed PBS.
4. Visualization: Observe immediately using a fluorescent microscope (Excitation ≈ 488 nm, Green emission for DNA, Red/Orange for RNA/acidic vesicles).

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

In Vivo

Acridine Orange (0.1 mg/kg, i.v., dogs) hydrochloride 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.

Molecular Weight

301.82

Formula

C₁₇H₂₀ClN₃

CAS No.

65-61-2

Appearance

Solid

Color

Brown to red

SMILES

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

Shipping

Room temperature in continental US; may vary elsewhere.

Storage

4°C, protect from light, stored under nitrogen

*In solvent : -80°C, 2 years; -20°C, 1 year (protect from light, stored under nitrogen)

Solvent & Solubility

In Vitro:

H₂O : 50 mg/mL (165.66 mM; ultrasonic and heat to 60°C)

DMSO : 25 mg/mL (82.83 mM; Need ultrasonic; Hygroscopic DMSO has a significant impact on the solubility of product, please use newly opened DMSO)

Preparing
Stock Solutions

Concentration Solvent Mass	1 mg	5 mg	10 mg

1 mM	3.3132 mL	16.5662 mL	33.1323 mL
5 mM	0.6626 mL	3.3132 mL	6.6265 mL
10 mM	0.3313 mL	1.6566 mL	3.3132 mL

View the Complete Stock Solution Preparation Table

* 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, 2 years; -20°C, 1 year (protect from light, stored under nitrogen). When stored at -80°C, please use it within 2 years. When stored at -20°C, please use it within 1 year.

* 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.

Molarity Calculator
Dilution Calculator

Mass (g) = Concentration (mol/L) × Volume (L) × Molecular Weight (g/mol)

Concentration _(start) × Volume _(start) = Concentration _(final) × Volume _(final)

This equation is commonly abbreviated as: C₁V₁ = C₂V₂

Concentration _(start)

Volume _(start)

Concentration _(final)

Volume _(final)

In Vivo Dissolution Calculator

Please enter the basic information of animal experiments:

Dosage

mg/kg

Animal weight
(per animal)

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(per animal)

μL

Number of animals

Recommended: Prepare an additional quantity of animals to account for potential losses during experiments.

Calculation results:

Working solution concentration: mg/mL

This product has good water solubility, please refer to the measured solubility data in water/PBS/Saline for details.

The concentration of the stock solution you require exceeds the measured solubility. The following solution is for reference only.If necessary, please contact MedChemExpress (MCE).

Purity & Documentation

Purity: 99.86%

Select Batch:

Data Sheet (275 KB) SDS (418 KB)

COA (246 KB) HNMR (180 KB) LCMS (340 KB)

Handling Instructions (2659 KB)

Dyeing Example

References

[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]

Cell Assay

Two-step, pH 3.0: Aliquots (0.2 mL, containing approximately 2-5x10⁵ 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 2x10^-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 4x10^-5 M^[2].

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

References

[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]

Complete Stock Solution Preparation Table

Optional Solvent	Concentration Solvent Mass	1 mg	5 mg	10 mg	25 mg

DMSO / H₂O	1 mM	3.3132 mL	16.5662 mL	33.1323 mL	82.8308 mL
	5 mM	0.6626 mL	3.3132 mL	6.6265 mL	16.5662 mL
	10 mM	0.3313 mL	1.6566 mL	3.3132 mL	8.2831 mL
	15 mM	0.2209 mL	1.1044 mL	2.2088 mL	5.5221 mL
	20 mM	0.1657 mL	0.8283 mL	1.6566 mL	4.1415 mL
	25 mM	0.1325 mL	0.6626 mL	1.3253 mL	3.3132 mL
	30 mM	0.1104 mL	0.5522 mL	1.1044 mL	2.7610 mL
	40 mM	0.0828 mL	0.4142 mL	0.8283 mL	2.0708 mL
	50 mM	0.0663 mL	0.3313 mL	0.6626 mL	1.6566 mL
	60 mM	0.0552 mL	0.2761 mL	0.5522 mL	1.3805 mL
	80 mM	0.0414 mL	0.2071 mL	0.4142 mL	1.0354 mL
H₂O	100 mM	0.0331 mL	0.1657 mL	0.3313 mL	0.8283 mL

* 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.