Aldh1 fitc

Expression of human NSCLC stem cell marker CD133 and CD44 was analyzed by flow cytometric study in A549 cells and spheroids by using anti-CD133-FITC and anti-CD44-APC antibodies (BD Biosciences). CD133⁺ and CD44⁺ CSCs were flow-cytometrically gated from spheroids on the basis of the cell surface phenotype. Mean fluorescence intensities of Oct-4-PerCP-Cy5.5, Nanog-PE, Sox-2- Alexa Fluor-647, Mrp1-FITC, Aldh1-FITC (BD Biosciences) were quantified^{22 (link)}. Mean fluorescence intensities of Akt, p-Akt, Cxcr-4, Mdr1, Integrin-α2, Integrin-α5, Integrin-β1, p-Fak (Santa Cruz Biotechnology, Inc.) were determined with respective primary antibodies conjugated with PE as previously described^{13 (link)}.

Expression of human bCSC markers CD44 and CD24 were analyzed by flow cytometric study in different stages of breast cancer tissue as well as in MCF-7/T47D cells and primary and secondary mammospheres by using CD44-FITC and CD24-PE antibodies (BD Biosciences). bCSCs were flow-cytometrically sorted from primary breast tumors on the basis of the cell surface phenotype CD44⁺/CD24^-/low. De-differentiation, drug resistance, and stemness phenomena were quantified flow-cytometrically by measuring mean fluorescence intensities of de-differentiation markers Oct-4-PerCP-Cy5.5, Nanog-PE, and Sox-2-Alexa Fluor-647; drug-resistance markers MRP1-FITC, ABCG2-PE, and ALDH1-FITC (BD Biosciences); and epithelial markers cytokeratin-18-PE and cytokeratin-19-PE (Santa Cruz Biotechnology, Inc., Santa Cruz, CA, USA). Expression levels of E-cadherin, β-catenin, and Slug (Santa Cruz Biotechnology, Inc.) were determined with respective primary antibodies conjugated with PE as previously described
[23 (link)].