Anti mouse cd133 pe antibody

Cells were dissociated from the dish using TE buffer (Invitrogen, Carlsbad, CA, USA) and labeled with an anti-mouse CD133-PE antibody (eBioscience, San Diego, CA, USA) on ice for 30 min following the standard protocol. The cells were analyzed using a FACS Calibur flow cytometer (San Jose, CA, USA). Summit software (FlowJo, Los Angeles, CA, USA) was used to determine the number of positive cells.

The CT26, CMT93 and RAW264.7 cells were harvested from the plates using PBS buffer (Invitrogen) and stained with an anti‐mouse CD133‐PE antibody (eBioscience, San Diego, CA). The tumour tissue was cut out from the rectum of mice and digested with fluid DNA enzyme I (ROCHE, 101041590001) and collagenase IV (Thermo Fisher, Carlsbad, CA, 17104019) for 1 hour, filter through 70‐μm filter, and additionally lysed with ACK Lysis Buffer (Solarbio, 20190726) for 5 minutes. Stained the dead cells (Thermo Fisher, L34964) and add the sealing solution (anti‐CD16/32, BioLegend, San Diego, CA, 101302), antibodies (anti‐CD11b, BD, 557396; anti‐CD45, BD, 560510; anti‐mouse F4/80, BioLegend, 123110), respectively. Then, the stained cells were analysed using a FACSCalibur flow cytometer (San Jose, CA).