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The cell cycle is of key importance to many areas of drug discovery. On the one hand this fundamental process provides the opportunity to discover new therapeutic targets for anti-cancer agents, and on the other hand drugs and targets in other therapeutic areas must be tested for undesirable effects on the cell cycle.
The Cell Proliferation Fluorescence Kit is a precise, fast, and simple fluorescence assay to quantitate cell proliferation, study the regulation of DNA synthesis, or observe the degree of inhibition of cell progression through S phase. It is based on the measurement of 5-bromo-2'-deoxyuridine (BrdU) incorporation during DNA synthesis in proliferating cells.
BrdU (an analog of the DNA precursor thymidine) is incorporated into newly synthesized DNA by cells entering and progressing through the S (DNA synthesis) phase of the cell cycle. The incorporated BrdU is detected with a specific anti-BrdU monoclonal antibody, followed by a Cy™5 labeled antibody to mouse immunoglobulin, giving a fluorescent signal at sites of BrdU incorporation. This provides an excellent marker for identifying cells that are in S phase, and also for determining the proportion of cells in S phase of the cell cycle.
The intensity of nuclear-associated Cy5 fluorescence is measured using an IN Cell Analyzer 1000 and the data analyzed using an Object Intensity Analysis Module.
The use of a Cy5 red-shifted fluor enables this technology to be multiplexed with green fluorescent protein (GFP). This assay provides a high-throughput method for the identification and analysis of cells that have actively synthesized DNA.
Materials
Products used
IN Cell Analyzer
1000
25-8010-26
Object Intensity Analysis Module 25-8010-56
for IN Cell Analyzer 1
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Source:
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