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 approximately 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).
Malanga
M
M
University of Zürich-Tierspital, Institute of Pharmacology and Toxicology, Switzerland.
Bachmann
S
S
Panzeter
P L
PL
Zweifel
B
B
Althaus
F R
FR
D000246:Adenosine Diphosphate Ribose / Q000378:metabolism*; D001704:Biopolymers; D002460:Cell Line; D006026:Glycoside Hydrolases; D006801:Humans; D008832:Microchemistry; D012015:Reference Standards; D012680:Sensitivity and Specificity; D016622:Silver Staining / Q000379:methods*
D001704:Biopolymers; D000246:Adenosine Diphosphate Ribose; D006026:Glycoside Hydrolases; C022407:poly ADP-ribose glycohydrolase