Detection and quantification of poly-ADP-ribosylated cellular proteins of spleen and liver tissues of mice in vivo by slot and Western blot immunoprobing using polyclonal antibody against mouse ADP-ribose polymer.
Poly-ADP-ribosylation (PAR) of cellular proteins has been shown to have decisive roles in diverse cellular functions including carcinogenesis. There are indications that metabolic level of poly-ADP-ribosylated cellular proteins might indicate carcinogenesis and, therefore, could be potentially used in cancer screening program. Keeping in mind the limitations of currently available assays of cellular PAR, a new assay is being reported that measures the metabolic level of poly-ADP-ribosylated cellular proteins. The ELISA based slot and Western blot immunoassay used polyclonal antibody against natural, heterogeneous ADP-ribose polymers. It could be successfully employed to qualitatively and quantitatively assay metabolic levels of poly-ADP-ribosylated proteins of spleen and liver tissues of normal mice or mice exposed to dimethylnitrosamine for up to 8 weeks; potentially PAR of cellular proteins could be assayed in any tissue or biopsy. Implications of the results in cancer screening program have been discussed.
Sharan
R N
RN
Radiation and Molecular Biology Unit, Department of Biochemistry, North-Eastern Hill University, Umshing, Shillong, India. rnsharan@nehu.ac.in
Devi
B Jaylata
BJ
Humtsoe
J O
JO
Saikia
Jyoti R
JR
Kma
L
L
D000246:Adenosine Diphosphate Ribose / Q000276:immunology*; D000818:Animals; D000906:Antibodies / Q000032:analysis* / Q000378:metabolism; D015153:Blotting, Western / Q000379:methods*; D004128:Dimethylnitrosamine / Q000378:metabolism / Q000494:pharmacology; D006657:Histones / Q000187:drug effects / Q000378:metabolism; D008099:Liver / Q000737:chemistry* / Q000201:enzymology; D051379:Mice; D008807:Mice, Inbred BALB C; D011064:Poly Adenosine Diphosphate Ribose / Q000032:analysis* / Q000737:chemistry; D011506:Proteins / Q000032:analysis* / Q000737:chemistry; D013154:Spleen / Q000737:chemistry* / Q000201:enzymology; D013997:Time Factors
D000906:Antibodies; D006657:Histones; D011506:Proteins; C024857:poly(ADP)-ribosylated proteins; D000246:Adenosine Diphosphate Ribose; D011064:Poly Adenosine Diphosphate Ribose; D004128:Dimethylnitrosamine