Acridine Orange hydrochloride

Name: Acridine Orange hydrochloride
Brand: MedChemExpress
SKU: HY-101879
Rating: 5.0 (21 reviews)

Cat. No.: HY-101879 Purity: 99.65%: FAQs
Data Sheet SDS COA Handling Instructions

Molarity Calculator

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.

Acridine Orange hydrochloride Chemical Structure

CAS No. : 65-61-2

Size	Price	Stock	Quantity

Free Sample (0.1 - 0.5 mg)		Apply Now
Solid + Solvent (Highly Recommended)
10 mM * 1 mL in DMSO ready for reconstitution	USD 66	In-stock	Estimated Time of Arrival: December 31
Solution
10 mM * 1 mL in DMSO	USD 66	In-stock	Estimated Time of Arrival: December 31
Solid
100 mg	USD 60	In-stock	Estimated Time of Arrival: December 31
200 mg		Get quote
500 mg		Get quote

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

Customer Review

Based on 21 publication(s) in Google Scholar

Other Forms of Acridine Orange hydrochloride:

Acridine Orange zinc chloride salt Get quote
Acridine Orange base Get quote

20 Publications Citing Use of MCE Acridine Orange hydrochloride

: 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

Guidelines (Following is our recommended protocol. This protocol only provides a guideline, and should be modified according to your specific needs)^[4].
1. Stain cells with Acridine Orange hydrochloride (1 μM; 20 min; 37℃).
2. Wash cells with PBS.
3. Cells are observed by a confocal laser scanning microscopy (FV3000, Olympus).

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

301.81

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 (165.67 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	3.3133 mL	16.5667 mL	33.1334 mL
5 mM	0.6627 mL	3.3133 mL	6.6267 mL
10 mM	0.3313 mL	1.6567 mL	3.3133 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:

Select the appropriate dissolution method based on your experimental animal and administration route.

For the following dissolution methods, please ensure to first prepare a clear stock solution using an In Vitro approach and then sequentially add co-solvents:
To ensure reliable experimental results, the clarified stock solution can be appropriately stored based on storage conditions. As for the working solution for in vivo experiments, it is recommended to prepare freshly and use it on the same day.
The percentages shown for the solvents indicate their volumetric ratio in the final prepared solution. If precipitation or phase separation occurs during preparation, heat and/or sonication can be used to aid dissolution.

Protocol 1

Add each solvent one by one: 10% DMSO 40% PEG300 5% Tween-80 45% Saline
Solubility: 2.08 mg/mL (6.89 mM); Suspended solution; Need ultrasonic

This protocol yields a suspended solution of 2.08 mg/mL. Suspended solution can be used for oral and intraperitoneal injection.
Taking 1 mL working solution as an example, add 100 μL DMSO stock solution (20.8 mg/mL) to 400 μL PEG300, and mix evenly; then add 50 μL Tween-80 and mix evenly; then add 450 μL Saline to adjust the volume to 1 mL.
Preparation of Saline: Dissolve 0.9 g sodium chloride in ddH₂O and dilute to 100 mL to obtain a clear Saline solution.
Protocol 2

Add each solvent one by one: 10% DMSO 90% (20% SBE-β-CD in Saline)
Solubility: ≥ 2.08 mg/mL (6.89 mM); Clear solution

This protocol yields a clear solution of ≥ 2.08 mg/mL (saturation unknown).
Taking 1 mL working solution as an example, add 100 μL DMSO stock solution (20.8 mg/mL) to 900 μL 20% SBE-β-CD in Saline, and mix evenly.
Preparation of 20% SBE-β-CD in Saline (4°C, storage for one week): 2 g SBE-β-CD powder is dissolved in 10 mL Saline, completely dissolve until clear.

For the following dissolution methods, please prepare the working solution directly. It is recommended to prepare fresh solutions and use them promptly within a short period of time.
The percentages shown for the solvents indicate their volumetric ratio in the final prepared solution. If precipitation or phase separation occurs during preparation, heat and/or sonication can be used to aid dissolution.

Protocol 1

Add each solvent one by one: PBS
Solubility: 10 mg/mL (33.13 mM); Clear solution; Need ultrasonic

In Vivo Dissolution Calculator

Please enter the basic information of animal experiments:

Dosage

mg/kg

Animal weight
(per animal)

Dosing volume
(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 (287 KB) SDS (393 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	3.3133 mL	16.5667 mL	33.1334 mL	82.8336 mL
	5 mM	0.6627 mL	3.3133 mL	6.6267 mL	16.5667 mL
	10 mM	0.3313 mL	1.6567 mL	3.3133 mL	8.2834 mL
	15 mM	0.2209 mL	1.1044 mL	2.2089 mL	5.5222 mL
	20 mM	0.1657 mL	0.8283 mL	1.6567 mL	4.1417 mL
	25 mM	0.1325 mL	0.6627 mL	1.3253 mL	3.3133 mL
	30 mM	0.1104 mL	0.5522 mL	1.1044 mL	2.7611 mL
	40 mM	0.0828 mL	0.4142 mL	0.8283 mL	2.0708 mL
	50 mM	0.0663 mL	0.3313 mL	0.6627 mL	1.6567 mL
	60 mM	0.0552 mL	0.2761 mL	0.5522 mL	1.3806 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.