Nonradioactive northern blotting for the determination of acetylcholinesterase mRNA. Comparison to the radioactive technique.
A sensitive nonradioactive northern blotting for the detection of acetylcholinesterase mRNA in mammalian tissues is described and compared to its radioactive version. Best results were obtained if digoxigenin labeled RNA probe was used for hybridization and CDP-Star, a chemiluminescent alkaline phosphatase substrate, for detection. The described nonradioactive technique for acetylcholinesterase mRNA determination is as sensitive as the radioactive one, but requires no protection against radiation and is less time consuming. Because of higher stability of the labeled probe, nonradioactive technique is also more convenient from the standpoint of experimental planning.
Zajc Kreft
K
K
Institute of Pathophysiology, School of Medicine, Ljubljana, Slovenia.
Kreft
S
S
Komel
R
R
Grubic
Z
Z
D000110:Acetylcholinesterase / Q000235:genetics*; D000818:Animals; D015152:Blotting, Northern / Q000379:methods*; D015335:Molecular Probes; D018482:Muscle, Skeletal / Q000201:enzymology; D012333:RNA, Messenger / Q000032:analysis*; D011868:Radioisotopes; D051381:Rats; D012680:Sensitivity and Specificity
D015335:Molecular Probes; D012333:RNA, Messenger; D011868:Radioisotopes; D000110:Acetylcholinesterase