Mouse anti tn igm mab

To analyse the expression of Tn antigen in Cosmc‐overexpressing cells and control cells, flow cytometry analysis was performed. Briefly, 1 × 10⁵ cells were routinely trypsinized, suspended in cold PBS and then incubated with 10 μg/mL mouse anti‐Tn IgM mAb or mouse IgM isotype‐antibody as a control (Santa Cruz, USA) for 2 hours at 4°C.4 After being washed three times with PBS, cells were incubated with PE‐labelled goat antimouse IgM secondary antibody (BD, USA) for 1 hour at 4°C. Cells were washed three times and suspended in 500 μL cold PBS per tube, followed by further analysis on a Flow Cytometer (Canto II, BD Bioscience, USA).

The expression of Tn antigen after T-synthase knockout in cells was analyzed by a Flow Cytometer (Canto II, BD Bioscience). In brief, 1×10⁵ cells were suspended in cold PBS and incubated with 10 μg/mL mouse anti-Tn IgM mAb (kindly provided by Dr. Tongzhong Ju, Emory University, Atlanta) or mouse IgM isotype-antibody as a control (Santa Cruz), for 2 hours at 4°C. Then the cells were washed with cold PBS three times and incubated with PE-labeled goat anti-mouse IgM secondary antibody (BD, 562033) for 1 hour at 4°C. After washing with PBS three times, the cells were analyzed by flow cytometry (Canto II, BD Bioscience).