Proliferation Assay: [3H] Thymidine incorporation

Proliferation Assay: [3H] Thymidine incorporationContributor: Suprya JayadevDate: September 13, 1994Labelling: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.