Flow cytometric analysis of RNA synthesis by detection of bromouridine incorporation
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Flow cytometric analysis of RNA synthesis by detection of bromouridine incorporation. / Larsen, J K; Jensen, Peter Østrup; Larsen, J.
In: Current protocols in cytometry / editorial board, J. Paul Robinson, managing editor ... [et al.], Vol. Chapter 7, 2001, p. Unit 7.12.Research output: Contribution to journal › Journal article › Research › peer-review
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TY - JOUR
T1 - Flow cytometric analysis of RNA synthesis by detection of bromouridine incorporation
AU - Larsen, J K
AU - Jensen, Peter Østrup
AU - Larsen, J
PY - 2001
Y1 - 2001
N2 - 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.
AB - 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.
KW - Animals
KW - Antigens
KW - Cell Membrane
KW - Cell Nucleus
KW - Cell Separation
KW - Flow Cytometry
KW - Gene Expression
KW - Humans
KW - RNA
KW - Time Factors
KW - Uridine
U2 - 10.1002/0471142956.cy0712s12
DO - 10.1002/0471142956.cy0712s12
M3 - Journal article
C2 - 18770724
VL - Chapter 7
SP - Unit 7.12
JO - Current Protocols in Cytometry
JF - Current Protocols in Cytometry
SN - 1934-9297
ER -
ID: 38517997