A simple and rapid method for the detection of poly(ADP-ribose) by flow cytometry.
Measurement of poly(ADP-ribose) levels was performed by a new method using a monoclonal antibody against poly(ADP-ribose) and flow cytometry from small amount of cultured cells without the need for isolation of poly(ADP-ribose) polymer. The increase of poly(ADP-ribose)-associated fluorescence intensity was observed in individual human leukemic HL-60 cells when treated with the carcinogen, N-methyl-N'-nitro-N-nitrosoguanidine (MNNG), and was blocked by the treatment with 3-aminobenzamide before MNNG treatment. It is easy and rapid to detect the time-dependent change of poly(ADP-ribose) levels in HL-60 cells after MNNG treatment. We easily found that the increase of the poly(ADP-ribose) level in nicotinic acid-treated lymphocytes after MNNG treatment was observed, but not in nicotinamide-treated lymphocytes. We investigated the change of poly(ADP-ribose) levels especially in the early phase of apoptosis. Our method is simple and rapid. It is suggested that the investigation of poly(ADP-ribosyl)ation in various fields is possible by using this new method.
Ogata
S
S
Laboratory of Biological Chemistry, Faculty of Bioresources, Mie University, Tsu, Japan.
Okumura
K
K
Taguchi
H
H
D001549:Benzamides / Q000494:pharmacology; D004791:Enzyme Inhibitors / Q000494:pharmacology; D005434:Flow Cytometry / Q000379:methods*; D018922:HL-60 Cells; D006801:Humans; D008769:Methylnitronitrosoguanidine / Q000494:pharmacology; D011064:Poly Adenosine Diphosphate Ribose / Q000032:analysis*; D000067856:Poly(ADP-ribose) Polymerase Inhibitors
D001549:Benzamides; D004791:Enzyme Inhibitors; D000067856:Poly(ADP-ribose) Polymerase Inhibitors; D008769:Methylnitronitrosoguanidine; D011064:Poly Adenosine Diphosphate Ribose; C025160:3-aminobenzamide