Detection of cyclin messenger RNAs by nonradioactive ribonuclease protection assay: a comparison of four detection methods.
Stable, nonradioactive riboprobes were used in ribonuclease protection assays to monitor the changes in cyclin mRNA expression during cell cycle progression in human mammary epithelial cells. Probes labeled with biotin demonstrated sufficient sensitivity (comparable to 32P) to detect these low-abundance mRNAs and thus offer a safe and easy alternative to traditional radioactive assays. Three detection systems based on chemiluminescence generated by horseradish peroxidase or alkaline phosphatase were compared for sensitivity, background and ease of use.
Nass
S J
SJ
Lombardi Cancer Center, Georgetown University, Washington, DC 20007, USA.
Dickson
R B
RB
D001426:Bacterial Proteins; D001710:Biotin; D001940:Breast; D002461:Cell Line, Transformed; D016213:Cyclins / Q000235:genetics*; D004848:Epithelium; D015870:Gene Expression; D006801:Humans; D008163:Luminescent Measurements; D009195:Peroxidase; D012333:RNA, Messenger / Q000032:analysis*; D012260:Ribonucleases; D019809:Streptavidin
D001426:Bacterial Proteins; D016213:Cyclins; D012333:RNA, Messenger; D001710:Biotin; D019809:Streptavidin; D009195:Peroxidase; D012260:Ribonucleases