Rat igg1 κ isotype control fitc

Cultured cells were detached with accutase solution (Sigma-Aldrich, Inc., St. Louis, MO, USA) and washed in phosphate-buffered saline (PBS) containing 0.5% FBS. Single cells were stained for 20 min on ice in the dark, washed twice in PBS containing 0.5% FBS, and fixed in 2% paraformaldehyde. Flow cytometry analysis was performed using a FACSCalibur system (BS Biosciences, San Jose, CA), and cell sorting was performed using FACSAria II (BD Immunocytochemistry System, Franklin Lakes, NJ). Antibodies against CD24 (anti-CD24-PE, BD), CD44 (anti-CD44-APC, BD), and ESA (anti-ESA-FITC, BD) were used. FITC-mouse IgG2b, κ isotype control (BD), rat IgG1 κ isotype control FITC (eBioscience), PE-mouse IgG2a, κ isotype control (BD), and APC-mouse IgG2b, κ isotype control (BD) were used as controls.

Cultured cells were detached using Accutase solution (Sigma Aldrich, Sigma-Aldrich, Inc., St. Louis, MO, US) and were washed in PBS with 0.5% FBS. Single cells were stained for 20 min on ice in the dark, washed twice in PBS with 0.5% FBS, and then fixed in 2% paraformaldehyde. Flow cytometric analysis was performed on a FACSCalibur system (BS Biosciences, San Jose, CA, US), and cell sorting was performed using FACSAria II (BD Immunocytochemistry System, Franklin Lakes, NJ, US). Antibodies against CD44 (anti-CD44-FITC, BD Pharmingen, Franklin Lakes, USA) and c-Met (anti-c-Met-FITC, eBioscience, San Diego, California, US) were used. Antibodies for cell sorting against CD24 (anti-CD24-PE, BD), CD44 (anti-CD44-APC, BD), and ESA (anti-ESA-FITC, BD) were used. FITC-mouse IgG2b, κ isotype control (BD), rat IgG1 κ isotype control FITC (eBioscience, San Diego, California, US), PE-mouse IgG2a, κ isotype control (BD Immunocytochemistry System, Franklin Lakes, NJ, US), and APC-mouse IgG2b, κ isotype control (BD Immunocytochemistry System, Franklin Lakes, NJ, US) were used as the controls.