Ivis spectrum imaging system

Seed MDA-MB-468 cells in a 96-well plate (density: 1.0 × 10⁴) in 100 μL complete L-15 medium. After overnight cultivation, switch to serum-free medium containing mAb_Nectin-4-ICG/free ICG with different ICG doses (1, 5, 10, 20, 30, 40, 50, 100, and 200 μg/mL) and continue incubating for 4 h. Rinsed the cells by PBS twice, and co-incubate the cells with 1 × Cell counting kit-8 (CCK-8) solution for another 4 h. Utilizing a multi-functional microplate reader to detect the absorbance at 450 nm and calculate the cell viability (n = 5).
Following same seeding conditions, after complete attachment, co-incubate MDA-MB-468 cells with mAb_Nectin-4-ICG or free ICG (20 μg/mL, 100 μL/well; within serum-free medium) for 4 h. The treated cells were flushed using PBS twice, then imaged with an IVIS Spectrum imaging system (Bruker, Germany; excitation/emission of 750/790 nm fliters). For blocking studies, cells were incubated previously with excess mAb_Nectin-4 for 1 h.

Seed MDA-MB-468 cells in a 96-well plate (density: 1.0 × 10⁴) in 100 μL complete L-15 medium. After overnight cultivation, switch to serum-free medium containing mAb_Nectin-4-ICG/free ICG with different ICG doses (1, 5, 10, 20, 30, 40, 50, 100, and 200 μg/mL) and continue incubating for 4 h. Rinsed the cells by PBS twice, and co-incubate the cells with 1 × Cell counting kit-8 (CCK-8) solution for another 4 h. Utilizing a multi-functional microplate reader to detect the absorbance at 450 nm and calculate the cell viability (n = 5).
Following same seeding conditions, after complete attachment, co-incubate MDA-MB-468 cells with mAb_Nectin-4-ICG or free ICG (20 μg/mL, 100 μL/well; within serum-free medium) for 4 h. The treated cells were flushed using PBS twice, then imaged with an IVIS Spectrum imaging system (Bruker, Germany; excitation/emission of 750/790 nm fliters). For blocking studies, cells were incubated previously with excess mAb_Nectin-4 for 1 h.

Seed MDA-MB-468 cells in a 96-well plate (density: 1.0 × 10⁴) in 100 μL complete L-15 medium. After overnight cultivation, switch to serum-free medium containing mAb_Nectin-4-ICG/free ICG with different ICG doses (1, 5, 10, 20, 30, 40, 50, 100, and 200 μg/mL) and continue incubating for 4 h. Rinsed the cells by PBS twice, and co-incubate the cells with 1 × Cell counting kit-8 (CCK-8) solution for another 4 h. Utilizing a multi-functional microplate reader to detect the absorbance at 450 nm and calculate the cell viability (n = 5).
Following same seeding conditions, after complete attachment, co-incubate MDA-MB-468 cells with mAb_Nectin-4-ICG or free ICG (20 μg/mL, 100 μL/well; within serum-free medium) for 4 h. The treated cells were flushed using PBS twice, then imaged with an IVIS Spectrum imaging system (Bruker, Germany; excitation/emission of 750/790 nm fliters). For blocking studies, cells were incubated previously with excess mAb_Nectin-4 for 1 h.