Proliferation Assay: [3H] Thymidine incorporation
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<center> <h2> <font>Proliferation Assay: [3H] Thymidine incorporation</font></h2> </center>
Contributor: Suprya Jayadev
Date: September 13, 1994
Labelling:
1) Seed cells in 6 well plates ( in normal growth media) such that there are 5 X 104 cells per well.
2) Incubate cells 2 nights.
3) Remove media, wash cells one time with 1X PBS, and add serum free media.
4) Rest cells for 2-4 hours then treat.
5) Six-twenty four hours before cells are to be harvested, add 1 µCi/ml [3H]-TdR to each well.
Harvesting:
6) Wash cells 2 times with ice cold PBS.
7) Wash cells 2 times with 5% TCA.
8) Solubilize cells by adding 0.5 ml 10.25 N NaOH (pipet up and down to completely solubilize cells).
9) Collect 400 µl of the solubilized cell solution into 4 ml scintillation vials and count.
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