What is thymidine incorporation assay?

Assay Description The thymidine incorporation assay, the most common assay, utilizes a strategy wherein a radioactive nucleoside, 3H-thymidine, is incorporated into new strands of chromosomal DNA during mitotic cell division. The advantage of these incorporation assays is that they are direct measures of proliferation.

Why is radiolabeled thymidine used?

When cells are incubated with thymidine, they use the radiolabeled thymidine to synthesize DNA and incorporate it into their DNA backbone. So, thymidine is an excellent measure of DNA synthesis in cells that have undergone the S-Phase of cell replication.

What is 3H-thymidine used for?

Metabolic incorporation of 3H-thymidine into cellular DNA is a widely used protocol to monitor rates of DNA synthesis and cell proliferation.

How do you measure DNA synthesis?

Measurement of new DNA synthesis is, therefore, essentially synonymous with measurement of cell proliferation. Direct measurement generally involves the incorporation of a labeled nucleoside into genomic DNA. Examples include the tritiated thymidine ([3H]dT) and BrdU (bromodeoxyuridine) methods 1, 2.

What is the purpose of the thymidine incorporation assay?

Assay Description. The thymidine incorporation assay, the most common assay, utilizes a strategy wherein a radioactive nucleoside, 3H-thymidine, is incorporated into new strands of chromosomal DNA during mitotic cell division. A scintillation beta-counter is used to measure the radioactivity in DNA recovered from the cells in order to determine

How are thymidine and BrdU used to measure cell proliferation?

DNA repair is taking place). So, Thymidine and BrdU assays are really DNA replication assays and not perfect cell proliferation assays. But, for the most part, they are the gold-standard when looking for cell proliferation. Thick tissue sections?

When to use thymidine triphosphate vs.probe labeling?

Proliferation vs. probes: Thymidine and methyl thymidine are used for proliferation studies, while thymidine triphosphate is used for probe labeling, because triphosphate is unable to penetrate the cell membrane. Grow cells to subconfluence in complete medium. Grow cells at 37°C in a humidified atmosphere with 5% CO 2 in 162 cm 2 flasks.

What happens when thymidine and BrdU are not used?

Taking things with a grain of salt: Note that DNA replication can happen even when cells are not proliferating. For example, if you have damaged DNA (ie. DNA repair is taking place). So, Thymidine and BrdU assays are really DNA replication assays and not perfect cell proliferation assays.

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