Fitc and pe conjugated anti mouse igg

Cells were stained using a FITC-conjugated Annexin V apoptosis detection kit (BD Biosciences, Franklin Lakes, NJ, USA) according to the manufacturer’s protocol. For CD44⁺/CD24^- staining analysis, cells were incubated for 30 min at 4°C with FITC- and PE-conjugated anti-mouse IgG or FITC-conjugated anti-CD24 and PE-conjugated anti-CD44 antibodies (BD Biosciences). Cells were analyzed by flow cytometry using a Beckman Coulter Expo.

ALDH1 activity was determined using an Aldefluor assay kit (Stemcell Technologies, Vancouver, BC), as previously described [43 (link)]. Diethylamino-benzaldehyde (DEAB) was used to define the Aldefluor-positive population. For CD44/CD24 staining, cells were stained with FITC- and PE-conjugated anti-mouse IgG or FITC-conjugated anti-CD24 and PE-conjugated anti-CD44 antibodies (BD Biosciences) and analyzed with a BD LSRFortessa™ X-20 Cell Analyzer.

ALDH1 activity was assessed using the Aldefluor assay kit (Stemcell Technology, Vancouver, BC). Cells were incubated at 37℃ for 45 min in Aldefluor assay buffer containing the ALDH1 protein substrate BODIPY-aminoacetaldehyde (BAAA, 1 µM per 0.5×10⁶ cells). To establish the baseline for Aldefluor-positive populations in flow cytometry, a specific inhibitor of ALDH1, diethylamino-benzaldehyde (DEAB), was used at a concentration of 50 mM. For CD44^high/CD24^low and CD49f^high/CD24^high staining, cells were incubated at 4℃ for 30 min with FITC- and PE-conjugated anti-mouse IgG or FITC-conjugated anti-CD24 and PE-conjugated anti-CD44 or CD49f antibodies (BD Biosciences) and analyzed by flow cytometry.