Acridine Orange hydrochloride hydrate

SKU: A0005

Categories: Fluorescent Detection, Fluorescent Detection

Synonyms: 3,6-Bis(dimethylamino)acridine hydrochloride , Acridine Orange N , Acridine Orange NO , Acridine Orange NS , Basic Orange 14 , Basic Orange 3RN , C.I. 46005 , C.I. Basic Orange 14 , Rhoduline Orange NO , Sumitomo Acridine Orange NO , 1704465-79-1; 65-61-2
CAS-Number: C₁₇H₁₉N₃ · HCl · xH₂O
Molecular Formula: 301.81 (anhydrous basis)
Molecular Weight

Product Data
Handling
Safety Information
Details

Specifications

Purity: ≥98% (HPLC)
Identity: 1H-NMR
Appearance: Orange to red powder

Properties

UV / Vis: λmax 492 nm
Melting Point: 284-287 °C (lit.)
Solvents: water (20 mg/ml) or DMSO (20 mg/ml).

Long Term Storage: +4°C
Use / Stability: Stable for at least 2 years after receipt when stored at +4°C.
Handling Advice: Protect from light and moisture.
Short Term Storage: RT
Shipping: AMBIENT

Transportation: Not dangerous goods
Signal word: Warning
GHS Symbol: GHS08
Precautionary statements: P201 - P202 - P280 - P308 + P313 - P405 - P501
Hazard statements: H341

Description: Acridine Orange hydrochloride salt is a cell-permeable metachromatic fluorescent dye that stains DNA and RNA. It is used as a nucleic acid-selective fluorescent cationic dye useful for cell cycle determination and staining dead cells. Being cell-permeable, it interacts with DNA and RNA by intercalation or electrostatic attractions respectively. When bound to DNA, it is very similar spectrally to fluorescein, with an excitation maximum at 502nm and an emission maximum at 525nm (green). When acridine orange associates with RNA, the excitation maximum shifts to 460nm (blue), and the emission maximum shifts to 650nm (red). Acridine orange will also enter acidic compartments such as lysosomes where it becomes protonated and sequestered. At low pH (inside the organelles), it will emit an orange fluorescence (peak at 590nm) and for optimal endosome visualization a blue light excitation (475nm) is used. Thus, acridine orange can be used to visualize primary lysosomes and phagolysosomes that may include products of phagocytosis of apoptotic cells. The dye is often used in epifluorescence microscopy and flow cytometry. It allows for visual detection of nucleic acids on agarose and polyacrylamide gels, can be used for differentiation of dsDNA (green fluorescence) and ssDNA/RNA (red fluorescence) and as a vitality test for determination of living cells. Spectral data: λEx=502nm, λEm=525nm (green, double strands) / λEx=460nm, λEm=650nm (red, single strands) / λEx=475nm, λEm=590nm (orange, acidic conditions).
InChi: InChI=1S/C17H19N3/c1-19(2)14-7-5-12-9-13-6-8-15(20(3)4)11-17(13)18-16(12)10-14/h5-11H,1-4H3
References: (1) F.H. Kasten, Int. Rev. Cytol. 21, 141 (1967) (Review) , (2) J.F. Golden & S.S. West, J. Histochem. Cytochem. 22, 495 (1974) , (3) J.F. Golden, et al., J. Histochem. Cytochem. 27, 522 (1979) , (4) J.K. Frost, et al., J. Histochem. Cytochem. 27, 545 (1979) , (5) H.W. Tyrer, et al., J. Histochem. Cytochem. 27, 552 (1979) , (6) R.N. Paul, Stain Technol. 55, 195 (1980) , (7) S. Mirrett, Inf. Contr. Hosp. Epidemiol. 3, 250 (1982) (Review) , (8) Z. Darzynkiewicz, et al., Curr. Protoc. Cytom. Chapter 7, Unit 7.3 (2004) (Review) , (9) J. Han & K. burgess, Chem. Revs. 110, 2709 (2010) , (10) R.W. Sabnis, Handbook of Fluorescent Dyes and Probes (2015) , (11) M.P. Thome, et al., J. Cell Sci. 129, 4622 (2016)
InChi Key: VSTHNGLPHBTRMB-UHFFFAOYSA-N
Smiles: CN(C)C1=CC=C2C(N=C(C=C(N(C)C)C=C3)C3=C2)=C1.[H]Cl

Acridine Orange hydrochloride salt is a cell-permeable metachromatic fluorescent dye that stains DNA and RNA. It is used as a nucleic acid-selective fluorescent cationic dye useful for cell cycle determination and staining dead cells. Being cell-permeable, it interacts with DNA and RNA by intercalation or electrostatic attractions respectively. When bound to DNA, it is very similar spectrally to fluorescein, with an excitation maximum at 502nm and an emission maximum at 525nm (green). When acridine orange associates with RNA, the excitation maximum shifts to 460nm (blue), and the emission maximum shifts to 650nm (red). Acridine orange will also enter acidic compartments such as lysosomes where it becomes protonated and sequestered. At low pH (inside the organelles), it will emit an orange fluorescence (peak at 590nm) and for optimal endosome visualization a blue light excitation (475nm) is used. Thus, acridine orange can be used to visualize primary lysosomes and phagolysosomes that may include products of phagocytosis of apoptotic cells. The dye is often used in epifluorescence microscopy and flow cytometry. It allows for visual detection of nucleic acids on agarose and polyacrylamide gels, can be used for differentiation of dsDNA (green fluorescence) and ssDNA/RNA (red fluorescence) and as a vitality test for determination of living cells. Spectral data: λEx=502nm, λEm=525nm (green, double strands) / λEx=460nm, λEm=650nm (red, single strands) / λEx=475nm, λEm=590nm (orange, acidic conditions).

SKU: A0005 Categories: Fluorescent Detection, Fluorescent Detection

Additional information

Additional information

Synonyms	3,6-Bis(dimethylamino)acridine hydrochloride, Acridine Orange N, Acridine Orange NO, Acridine Orange NS, Basic Orange 14, Basic Orange 3RN, C.I. 46005, C.I. Basic Orange 14, Rhoduline Orange NO, Sumitomo Acridine Orange NO, 1704465-79-1
Purity	≥98% (HPLC)
Appearance	Orange to red powder
CAS-Number	65-61-2
Molecular Formula	C17H19N3 · HCl · xH2O
Molecular Weight	301.81 (anhydrous basis)
Identity	1H-NMR
Solvents	water (20 mg/ml) or DMSO (20 mg/ml).
Melting Point	284-287 °C (lit.)
UV / Vis	λmax 492 nm
Smiles	CN(C)C1=CC=C2C(N=C(C=C(N(C)C)C=C3)C3=C2)=C1.[H]Cl
InChi Key	VSTHNGLPHBTRMB-UHFFFAOYSA-N
Shipping	AMBIENT
Short Term Storage	RT
Long Term Storage	+4°C
Handling Advice	Protect from light and moisture.
Use / Stability	Stable for at least 2 years after receipt when stored at +4°C.
Hazard statements	H341
Precautionary statements	P201 – P202 – P280 – P308 + P313 – P405 – P501
GHS Symbol	GHS08
Signal word	Warning
Transportation	Not dangerous goods
References	(1) F.H. Kasten, Int. Rev. Cytol. 21, 141 (1967) (Review), (2) J.F. Golden & S.S. West, J. Histochem. Cytochem. 22, 495 (1974), (3) J.F. Golden, et al., J. Histochem. Cytochem. 27, 522 (1979), (4) J.K. Frost, et al., J. Histochem. Cytochem. 27, 545 (1979), (5) H.W. Tyrer, et al., J. Histochem. Cytochem. 27, 552 (1979), (6) R.N. Paul, Stain Technol. 55, 195 (1980), (7) S. Mirrett, Inf. Contr. Hosp. Epidemiol. 3, 250 (1982) (Review), (8) Z. Darzynkiewicz, et al., Curr. Protoc. Cytom. Chapter 7, Unit 7.3 (2004) (Review), (9) J. Han & K. burgess, Chem. Revs. 110, 2709 (2010), (10) R.W. Sabnis, Handbook of Fluorescent Dyes and Probes (2015), (11) M.P. Thome, et al., J. Cell Sci. 129, 4622 (2016)
InChi	InChI=1S/C17H19N3/c1-19(2)14-7-5-12-9-13-6-8-15(20(3)4)11-17(13)18-16(12)10-14/h5-11H,1-4H3
Quantity	1 g, 5 g, Bulk