Acridine Orange CAS 10127-02-3 Electrophoresis Grade / Bio-Tech Grade orange-red powder fluorescent dye / nucleic acid and cell stain fluorescence ind

1. Product Overview

Acridine Orange (also known as Basic Orange 14, C.I. 46005; CAS 10127-02-3) is a cationic fluorescent metachromatic dye and a zinc chloride double salt of 3,6-bis(dimethylamino)acridine. The commonly supplied form is C₁₇H₂₀Cl₃N₃Zn (MW 438.13) or the hemizinc chloride salt C₁₇H₂₀ClN₃·HCl·½ZnCl₂ (MW 369.96). It appears as an orange-red to brown powder, freely soluble in water and ethanol to give an orange-yellow solution with intense greenish fluorescence under UV/blue excitation (Ex ~488 nm, Em ~515–530 nm for DNA; orange-red for RNA). Supplied as Electrophoresis Grade (dye content ~80–90%, ≥80–90%, Em(490 nm, water) >36,500), it is widely used as a vital stain for nuclei acids, lysosomes, and acidic vesicles in fluorescence microscopy and flow cytometry, as well as a nucleic acid stain in agarose/polyacrylamide gel electrophoresis. Also functions as a frameshift mutagen and fluorescence indicator in biochemical research. For lab/research use only; not for human/veterinary therapeutic application.

2. Key Features and Advantages

• Metachromatic fluorescence: binds to dsDNA (green, ~525 nm) and ssDNA/RNA (orange-red, ~650 nm), enabling differential staining of nucleic acids in cells and gels.

• High quantum yield with strong greenish fluorescence in aqueous solution; excitation at 488 nm (argon laser) or 450–490 nm (blue filter), emission 515–650 nm depending on target.

• Electrophoresis Grade with dye content ~80–90% and low background; ideal for sensitive detection of nucleic acids in gels (detects as low as nanogram levels).

• Cell-permeant and vital stain properties: penetrates live cell membranes to stain lysosomes, acidic organelles, and apoptotic bodies (AO/EB dual staining for apoptosis assays).

• Stable as dry powder at RT; water-soluble (≥1 mg/mL, clear orange to deep orange solution), compatible with standard buffer systems (pH 6.0–7.0 optimal; loses fluorescence at pH 8.4–10.4).

• Available in CN reagent grade (BR/Bio-Tech ≥80%, Electrophoresis ≥90%) and international premium brands (Sigma A6014, TCI, Alfa Aesar).

3. Main Applications

• Fluorescence microscopy & flow cytometry: vital staining of nuclei acids in live/dead cells, lysosomal tracking, autophagy/acidic vesicle staining, AO/EB or AO/PI dual staining for apoptosis detection.

• Gel electrophoresis: nucleic acid stain for agarose and polyacrylamide gels (alternative to ethidium bromide, detects dsDNA/ssDNA/RNA simultaneously with metachromatic shift).

• Microbiology & cytology: staining of bacteria, tumor cells, blood smears; differentiation of viable vs non-viable cells based on membrane integrity and AO uptake.

• Mutagenesis research: used as a frameshift mutagen intercalating between DNA base pairs, causing insertion/deletion mutations in replication studies.

• Analytical chemistry: fluorescence indicator for pH (green fluorescence at acidic pH, quenched at alkaline), metal-ion sensing, and chromatographic visualization.

• Biochemical education & kit components: included in cell biology teaching kits and commercial apoptosis/viability assay kits.

4. Technical Specifications

5. Storage and Handling

Store as dry powder in a tightly sealed amber vial at 15–25 °C (room temperature), protected from light and moisture; for long-term (>12 months) keep at 2–8 °C in desiccator. This compound is hygroscopic and light-sensitive (fluorescence quenches under prolonged UV/visible light). Avoid exposure to strong oxidizing agents. Handle in a well-ventilated area with nitrile gloves, lab coat, and safety goggles; minimize dust generation. Prepare working solutions fresh in water or buffer (1 mg/mL stock, filter if needed); store solution at 2–8 °C protected from light and use within 1–2 weeks (degrades in solution). Do not eat, drink, or smoke in handling areas. Wash hands thoroughly after use. Decontaminate surfaces with bleach solution (hypochlorite breaks down acridine dyes). Dispose as hazardous mutagenic/organic waste per local regs (WGK Germany 3).

6. Safety Overview

Classified under GHS as Warning/Danger (GHS07/GHS08): Hazard statements H302 (Harmful if swallowed), H315 (Causes skin irritation), H319 (Causes serious eye irritation), H335 (May cause respiratory irritation), H341 (Suspected of causing genetic defects—frameshift mutagen). Precautionary: P201 (Obtain special instructions before use), P261 (Avoid breathing dust), P264 (Wash after handling), P271 (Use outdoors/well-ventilated), P280 (Protective gloves/eyewear), P305+P351+P338 (Eye rinse), P308+P313 (If exposed/suspected, get medical advice). Not for human/veterinary therapeutic use; for authorized lab research, microscopy, electrophoresis only. Mutagenic in Ames test; handle as potential carcinogen/mutagen. TCI ships as UN3077 Class 9 PG III (env. hazardous substance, solid, n.o.s.) in some regions. Consult full SDS before use; keep away from strong oxidizers and children.