Anti human cd133 apc

Peripheral blood samples were obtained from CRC patients attending our department and an informed consent was obtained from all the individuals. Peripheral blood samples were collected and prepared as per the protocol described in our previous report 19. In detail, CTCs from cell suspensions were characterized by multiparameter flow cytometry. The antibodies used in this study included anti‐human CD133‐APC, CD44‐FITC, CD54‐PErcp‐cy_5.5, CD54‐PE, and CD45‐BV₅₁₀ (all antibodies were purchased from BD Biosciences, San Diego, CA, USA). DAPI was used to identify and sort the dead cells. The remaining steps were the same as the protocol described in our previous report 19. The absolute CTCs or antibody‐positive cell numbers were derived from the absolute number of white blood cells provided by the hematological analyzer, and the percentage of CTCs or antibody‐positive cells was determined by flow cytometry, using the following formula: percentage of cells × white blood cells count/100.

CTCs from cells suspension were characterized by multiparameter flow cytometry. The antibodies used in this study include: anti-human CD133-APC, CD44-FITC, CD44-APC-Cy7, CD54-PErcp-cy5.5, CD54-PE, CD24-PE/Cy7, CD10-PECF594, CD26-PE, CD166-Percp-cy5.5, CD45-BV510, CD58-PE, CD66-PE, CD71-PE, CD117-PE, EPCAM-Percp-cy5.5, and EGFR-PE (all of the above-mentioned antibodies were purchased from BD Biosciences). DAPI was used to identify the dead cells. Evaluation of nucleated cells from whole cells suspensions was carried out using a FACS Canto Flow Cytometer (BD Biosciences) and data were analyzed using BD FACS Diva software. A range of internal quality assurance procedures was employed, including daily calibration of the optical alignment and fluidic stability of the flow cytometer using the seven-color Set-up Beads (BD Biosciences). The absolute CTCs or antibody-positive cell number was derived from the absolute number of the white blood cells provided by the hematological analyzer and percentage of CTCs or antibody-positive cell as determined by flow cytometry, using the following formula: percentage of cells × white blood cells count/100.