Flow cytometric analysis of RNA synthesis by detection of bromouridine incorporation.
RNA synthesis has traditionally been investigated by a laborious and time-consuming radiographic method involving incorporation of tritiated uridine. Now a faster non-radioactive alternative has emerged, based on immunocytochemical detection. This method utilizes the brominated RNA precursor bromouridine, which is taken into a cell, phosphorylated, and incorporated into nascent RNA. The BrU-substituted RNA is detected by permeabilizing the cells and staining with certain anti-BrdU antibodies. This dynamic approach yields information complementing that provided by cellular RNA content analysis at a given time and may be of value in studies of cellular activation and gene expression.
Larsen
J K
JK
The Finsen Laboratory, Rigshospitalet, Copenhagen, Denmark.
Jensen
P Ø
PØ
Larsen
J
J
D000818:Animals; D000941:Antigens / Q000096:biosynthesis; D001976:Bromouracil / Q000031:analogs & derivatives; D002462:Cell Membrane / Q000378:metabolism; D002467:Cell Nucleus / Q000378:metabolism; D002469:Cell Separation / Q000379:methods*; D005434:Flow Cytometry / Q000379:methods*; D015870:Gene Expression; D006801:Humans; D012313:RNA / Q000032:analysis*; D013997:Time Factors; D014529:Uridine / Q000031:analogs & derivatives* / Q000032:analysis / Q000494:pharmacology
D000941:Antigens; D001976:Bromouracil; D012313:RNA; C006824:5-bromouridine; D014529:Uridine