Comparison of various ultraviolet sources for fluorescent detection of ethidium bromide-DNA complexes in polyacrylamide gels

Absorption, fluroescence and circular dichroism measrements on 33258 Hoechst-deoxyribonucleic acid (DNA) complexes are consistent with the existence of two types of dye-binding interactions. One type, which persists at elevated solution ionic strength, is highly specific for adenine-thymine-rich DNA. Dye bound under this condition exhibits efficient fluorescence and strong optical activity. A less specific, largely electrostatic interaction is associated with less intense fluorescence and weaker optical activity. The fluorescence of 33258 Hoechst and several other bisbenzimidazole dyes is less when bound to poly(deoxyadenylate-5-bromodeoxyuridylate) than when bound to poly(deoxyadenlyate-deoxythymidylate). Quenching of 33258 Hoechst fluorescence can also be used to detect biosynthetic incorporation of 5-bromodeoxyuridine into the DNA of living cells. This property of 33258 Hoechst should allow fluorescence-activated cell and chromosome sorting according to the extent of DNA synthesis, providing a bridge between biochemical and cytologic analyses of processes related to DNA replication.

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Determination of DNase activity by degradation of ethidium bromide–DNA complexes using a fluorescence plate reader

Analytical Biochemistry ◽

10.1016/j.ab.2014.11.013 ◽

2015 ◽

Vol 471 ◽

pp. 73-79 ◽

Cited By ~ 2

Author(s):

Benjamin Vogel ◽

Stefan Frantz

Keyword(s):

Ethidium Bromide ◽

Dnase Activity ◽

Dna Complexes ◽

Plate Reader ◽

Fluorescence Plate Reader

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Rapid and Simple Nanogram Fluorescent Detection of Glycoproteins in Polyacrylamide Gels

The FASEB Journal ◽

10.1096/fasebj.21.5.a635-a ◽

2007 ◽

Vol 21 (5) ◽

Author(s):

Babu Siddegowda Antharavally ◽

Kenneth J Maas ◽

Krishna A Mallia ◽

Peter A Bell

Keyword(s):

Fluorescent Detection ◽

Polyacrylamide Gels

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Fast analysis of glycosaminoglycans by microchip electrophoresis with in situ fluorescent detection using ethidium bromide

Journal of Pharmaceutical and Biomedical Analysis ◽

10.1016/j.jpba.2004.11.024 ◽

2005 ◽

Vol 37 (3) ◽

pp. 429-436 ◽

Cited By ~ 9

Author(s):

Yu-ki Matsuno ◽

Mitsuhiro Kinoshita ◽

Kazuaki Kakehi

Keyword(s):

Ethidium Bromide ◽

Microchip Electrophoresis ◽

Fluorescent Detection ◽

Fast Analysis

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Subcellular distribution of an ethidium bromide/poly r(A-U) complex

Proceedings, annual meeting, Electron Microscopy Society of America ◽

10.1017/s0424820100146825 ◽

1993 ◽

Vol 51 ◽

pp. 196-197

Author(s):

J.M. Jamison ◽

J.L. Summers ◽

J. Gilloteaux

Keyword(s):

Ethidium Bromide ◽

Mitochondrial Swelling ◽

Drug Uptake ◽

Cell Multiplication ◽

Dna Complexes ◽

Viral Inactivation ◽

Double Stranded Rna ◽

Human Foreskin Fibroblasts ◽

Ehrlich Ascites ◽

Nucleolar Segregation

Previous results have shown that drug uptake by cells is enhanced when the drug is bound to DNA. Chick embryo fibroblasts (CEF) treated with ethidium bromide (EB) exhibit inhibited cell multiplication, mitochondrial swelling and nucleolar segregation. However, EB-DNA treated CEF cells exhibit a much lower cytotoxicity than the EB-treated CEF cells and the mitochondrial and nucleolar alterations are reversible. Conversely, in Ehrlich ascites tumor cells exposed to EB or EB-DNA complexes, the toxicity of the DNA bound EB (as measured by inhibition of cell multiplication, nucleolar segregation, mitochondrial swelling and intramitochondrial inclusions) is more pronounced and appears earlier after treatment than the EB induced cytotoxicity. More recent results have shown that when EB was combined with double stranded RNA (poly r(A-U)) and then added to human foreskin fibroblasts (HSF), the 50% effective dose of the poly r(A-U) was 154-fold lower. The results of additional studies demonstrated that the enhanced antiviral activity was not due to superinduction of interferon, direct viral inactivation, or host cell cytotoxicity.

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