Cd166 pe conjugated antibody

All cells were grown to the appropriate low and high culture densities and lifted using an (Ethylenedinitrilo)tetraacetic acid (EDTA) based solution called Cell Stripper (Fisher Scientific). Cells were spun for five minutes to pellet and resuspended in a 3% BSA blocking solution made in PBS. Cells were incubated for 1 hour on ice in this blocking solution before 5 µl of CD166-PE conjugated antibody (BD Biosciences) was added to the cells. An appropriate PE conjugated IgG control (BD Biosciences) was used at an equal concentration to the CD166 antibody. Cells were incubated on ice for 30 minutes. Following primary antibody and IgG incubation, cells were washed twice with PBS, and then resuspended in PBS. Cell suspensions were passed through a BD round bottom tube with a 50 µm cell strainer (BD Biosciences). Cell surface staining was analyzed using the BD FACSAria II Flow Cytometer.