Ab126556

Cell proliferation was examined using bromodeoxyuridine (BrdU) incorporation assay (Abcam, ab126556) according to manufacturer’s instructions.

Cell proliferation was assessed by alamarBlue staining (Thermo Scientific, DAL1100) or Bromodeoxyuridine (BrdU) incorporation assay (Abcam, ab126556).

To analyze differences in cellular proliferation, the BrdU assay (ab126556, Abcam) was performed for three replicates at D0, D5, and D28 time points. Twenty-four hours prior to the read-out, 1X-BrdU reagent was added to the cell culture vessels for incorporation by incubation at 37°C and 5% CO₂. The culture media was aspirated for the read-out, and cells were fixed with the supplied fixing solution. This was followed by exposure to the anti-BrdU antibody (primary antibody), incubation at room temperature for 1 h, and washing with the supplied plate wash buffer. The cells were incubated with the HRP-tagged secondary antibody at room temperature for 30 min, followed by TMB exposure and recording absorbance at 450 nm. Afterwards, the cells were incubated in TBS, 0.1% Triton-X 100, and Hoechst 1:1000 for 15 min, followed by fluorescence measurement, which was used to calibrate the BrdU quantification for the number of nuclei in each sample. The quantification of proliferation rate followed by Student's t-test statistical comparison was done in R (v4.1.2).

Cell proliferation was assayed by enzyme-linked immunosorbent assay (ELISA) using bromodeoxyuridine/5-bromo-2′-deoxyuridine (BrdU) labelling in accordance with manufacturer instructions (ab126556, Abcam). Cells were seeded at a density of 10,000 cells per well in a 96 well plate and incubated with BrdU for a period of 24 h following experimentation. Absorbance was then read at room temperature at 420 nm using a BioTek™ PowerWave™ Microplate Spectrophotometer.