Poly(ADP-ribose) Quantification at the Femtomole Level in Mammalian Cells

ADP-ribose polymers were isolated from living mammalian cells, separated by polyacrylamide gel electrophoresis, visualized in the gel with a novel silver staining agent, and quantified by computer-aided scanning densitometry. This method detects as little as ∼40 fmol of ADP-ribose polymers of a part...

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Bibliographic Details
Published inAnalytical biochemistry Vol. 228; no. 2; pp. 245 - 251
Main Authors Malanga, M., Bachmann, S., Panzeter, P.L., Zweifel, B., Althaus, F.R.
Format Journal Article
LanguageEnglish
Published United States Elsevier Inc 01.07.1995
Subjects
Adenosine Diphosphate Ribose - metabolism
Biopolymers
Cell Line
Glycoside Hydrolases
Humans
Microchemistry
Reference Standards
Sensitivity and Specificity
Silver Staining - methods
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Summary:ADP-ribose polymers were isolated from living mammalian cells, separated by polyacrylamide gel electrophoresis, visualized in the gel with a novel silver staining agent, and quantified by computer-aided scanning densitometry. This method detects as little as ∼40 fmol of ADP-ribose polymers of a particular size class and reduces the gel exposure times required for conventionally radiolabeled polymers from 2 months to about 1 h. The method also detects polymers with slow turnover which may be underestimated by techniques requiring metabolic radiolabeling of poly(ADP-ribose).
Bibliography:ObjectType-Article-1
SourceType-Scholarly Journals-1
ObjectType-Feature-2
content type line 23
ISSN:0003-2697
1096-0309
DOI:10.1006/abio.1995.1346