Aldefluor stem cell kit

Aldehyde dehydrogenase (ALDH) activity in DCA-treated cells was determined using an Aldefluor Stem Cell kit (StemCell Technologies, Vancouver, Canada). Cells were seeded into 6-well plates (100,000 cells/well) and treated with DCA (0.7 µM) for 48 h. Then, cells were incubated in 0.5 ml Aldefluor assay buffer containing ALDH substrate (5 µl/ml) for 45 min at 37 °C. As a negative control, cells were treated with 5 µl of diethylaminobenzaldehyde (DEAB; 50 mmol/l), a specific ALDH inhibitor. The percentage of ALDH-positive cells was determined by flow cytometry and analyzed using Flowing Software 2.5.1.

Aldehyde dehydrogenase (ALDH) activity was determined using an Aldefluor Stem Cell kit (StemCell Technologies, Vancouver, BC, Canada). The 4T1 cells were seeded on six-well plates (4T1, 100,000 cells/well) and treated with the indicated concentrations of IS for 24 h. Then, the collected cells were processed according to the manufacturer’s instructions. The SKBR-3 cell line was used for positive control samples based on the manufacturers’ instructions. Changes in the level of ALDH were assessed by flow cytometry and the results were analyzed using Flowing Software 2.5.1 (Beckton-Dickinson, Franklin Lakes, NJ, USA). The Aldefluor assay for assessing stemness was performed similarly to references [17 (link),100 (link),101 (link)].

Aldehyde dehydrogenase (ALDH) activity was measured using an Aldefluor Stem Cell kit (StemCell Technologies, Vancouver, BC, Canada). 4T1 cells were seeded on 6-well plates (4T1—100,000 cells/well) and treated with different concentrations of IPA for 24 h. Then, cells were collected and prepared according to the manufacturer’s instructions. For positive control samples, the SKBR-3 cell line was used based on the manufacturers’ instructions. Changes in the level of ALDH were determined by flow cytometry, and the results were analyzed by using Flowing Software 2.5.1. Aldefluor assay for assessing stemness was performed similarly to [28 (link),58 (link),59 (link)].