[cell biology work] Foreskin_fibroblast_proliferation_assay1

This topic has 0 replies, 1 voice, and was last updated 16 years, 6 months ago by Seung Jae Jeong.

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      Seung Jae Jeong
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      Foreskin fibroblast proliferation assay

       

      For thymidine incorporation assay, Cells(FF: 3X104)were seeded on 24 well plate and grown in DMEM medium supplemented with 10% fetal bovine serum and 50ug/ml streptomycin and penicillin in a 5% CO2 incubator at 37 degree for 12hrs. Culture medium was changed to serum free media with 1% streptomycin and penicillin without fetal bovine serum and incubated for 3hrs. p43 was treated as indicated concentration and cultured for 12hrs again. 3H labeled thymidine was treated with 1uci per well and incorporated to cells for 3 hrs. Cells were fixed with 5% TCA for 10 min and washed with 1X PBS 3 times. Cells were lysed by 0.5N NaOH and counted with Liquid scintillation counter.

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