The protein effect on determination of DNA with Hoechst 33258

The present study was designed to afford a critical review of the effect of proteins on the Hoechst 33258 method for determination of DNA of crude homogenates. A considerable effect of proteins on the fluorescence was observed when the concentration exceeded 100 μg BSA equivalent protein. Below that...

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Bibliographic Details
Published inJournal of biochemical and biophysical methods Vol. 28; no. 4; pp. 263 - 276
Main Authors Moe, D., Garbarsch, C., Kirkeby, S.
Format Journal Article
LanguageEnglish
Published Netherlands Elsevier B.V 01.06.1994
Subjects
Animals
Bisbenzimidazole
Deoxyribonucleases
DNA - analysis
DNA method
Endopeptidase K
Female
Fluorescence
Hoechst 33258
Kidney - chemistry
Liver - chemistry
Protein interface
Rats
Rats, Wistar
Reference Standards
Serine Endopeptidases
Serum Albumin, Bovine
Hoechst 33258
DNA method
Protein interface
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Summary:The present study was designed to afford a critical review of the effect of proteins on the Hoechst 33258 method for determination of DNA of crude homogenates. A considerable effect of proteins on the fluorescence was observed when the concentration exceeded 100 μg BSA equivalent protein. Below that value, practically no effect of proteins was noted. We used proteinase K to remove the proteins, but dilution of homogenates could be used as well. Moreover, we found that the concentration of the fluorochrome should be between 1 μg and μg when microgram levels DNA are to be determined.
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ISSN:0165-022X
1872-857X
DOI:10.1016/0165-022X(94)90002-7