Brdu incorporation elisa colorimetric assay

Cells (3×10³ per well) were seeded onto 96-well plates, after applied treatment, cell proliferation was assessed via the BrdU incorporation ELISA colorimetric assay (Roche, Indianapolis, IN) with the manufacturer’s protocol. The ELISA OD value of treatment group was normalized to that of untreated control group.

The HCC cells and hepatocytes were seeded initially (3×10³ per well, into 96-well tissue culture plate), and were exposed to JLGTE. After the indicated time period, BrdU incorporation ELISA colorimetric assay (Roche, Indianapolis, IN) was utilize to test cell proliferation. The ELISA OD at 450 nm was recorded.

Cell proliferation was evaluated by BrdU incorporation ELISA colorimetric assay (Roche Applied Science, Penzberg, Germany). In brief, cells were seeded in 96-well plates coated with poly-l-lysine (Sigma-Aldrich, St. Louis, MO, USA) at 5x10³/well and treated as described in the study design section. As a positive control, cells were treated with culture medium supplemented with 10% fetal bovine serum (FBS) [47 (link)]. Colchicine (1 μM) was used as a negative control, as previously reported [27 (link)]. At the end of the treatment period, cells were incubated with 10 μM bromodeoxyuridine (BrdU)-labelling solution per well for 4 h, dried, fixed and detected using an anti-BrdU antibody according to the manufacturer’s instructions. Finally, the absorbance was measured using a microplate reader at 450 nm with a reference wavelength of 690 nm.