Ethidium bromide 

Ethidium bromide (EtBr) is the most commonly used nucleic acid stain for PAGE or agarose gel electrophoresis and is available in several forms including a 10 mg/ml solution, 10 or 100 mg tablets or as a powder. The tablets and the solution help to decrease exposure to a potentially toxic chemical. Ethidium bromide intercalates double-stranded DNA and RNA. The fluorescence of EtBr increases 21-fold upon binding to double-stranded RNA and 25-fold on binding double-stranded DNA so that destaining the background is not necessary with a low stain concentration (10 μg/ml). Ethidium bromide has been used in a number of fluorimetric assays for nucleic acids. It has been shown to bind to single-stranded DNA (although not as strongly) and triple-stranded DNA.

Ethidium Bromide is a nucleic acid intercalating agent and frameshift mutagen. It can also be used in conjunction with acridine orange to differentiate between viable, apoptotic and necrotic cells.

For staining a gel after electrophoresis, dilute a sample of the stock solution to 0.5 μg/ml with water and incubate the gel for 15-30 min. Destaining is usually not needed but can be carried out in water for 15 min if decreased background is necessary. The DNA bands can then be detected on a UV light box (254 nm wavelength). Ethidium bromide can also be incorporated into the gel and running buffer at 0.5 μg/ml and visualized immediately after electrophoresis.

~95% (HPLC)

260-262 °C (dec.)(lit.)

Sambrook, J. Molecular Cloning: A Laboratory Manual Cold Spring Harbor, , (1989) 6-44, 6

Severini, A. , Morgan, A.R., An assay for proteinases and their inhibitors based on DNA/ethidium bromide fluorescence. Anal. Biochem. 193, 83, (1991)

Moore, S.P. , Sutherland, B.M., A densitometric nondestructive microassay for DNA quantitation. Anal. Biochem. 144, 15, (1985)

Karsten, U. , Wollenberger, A., Improvements in the ethidium bromide method for direct fluorometric estimation of DNA and RNA in cell and tissue homogenates. Anal. Biochem. 77, 464, (1977)

Bauer, W. , Vinograd, J., Interaction of closed circular DNA with intercalative dyes. II. The Free energy of superhelix formation in V40 DNA. Mol. Biol. 47, 410, (1970)

Gorman, A. Techniques in Apoptosis: A User’s Guide London, UK , (1996), 7

Scaria, P.V. , Shafer, R.H., Binding of ethidium bromide to a DNA triple helix. Evidence for intercalation. J. Biol. Chem. 266, 5417, (1991)

El-Hamalawi, A., The fluorometric determination of nucleic acids in pea seeds by use of ethidium bromide complexes. Anal. Biochem. 67, 384, (1975)

Merck 13,4751

Beil. 22,V,11,352

Aldrich MSDS 1, 864:C / Corp MSDS 1 (1), 1549:B / FT-IR 2 (3), 3935:C / FT-IR 1 (2), 873:B / IR-Spectra (3), 1399:A / IR-Spectra (2), 1220:B / RegBook 1 (2), 2635:A / Sax 6, 885 / Sigma FT-IR 1 (2), 136:D / Stains and Dyes Ref, 318 / Structure Index 1, 415:E:4

UN2811 6.1/PG 1

3

SF7950000