Anti-Human L1CAM Therapeutic Antibody Fab Fragment

Protocol tips

Upstream tips	Protocol tips	Downstream tips
	Cells were incubated with the human anti-L1CAM mAb, Ab417 (Cho et al., 2016), for 1 h at 4°C. After washing twice with phosphate buffered saline (PBS), cells were incubated with an anti-human IgG-FITC conjugate (Sigma) for 1 h at 4°C. After washing, propidium iodide (PI) was added to cells (Sigma), with PI-negative.	Cells were analyzed for bound antibody using a FACSCalibur flow cytometer (Becton-Dickinson, USA). Data acquisition and analyses were with the CellQuest software (Becton-Dickinson).

Protocol tips
Cells were incubated with the human anti-L1CAM mAb, Ab417 (Cho et al., 2016), for 1 h at 4°C. After washing twice with phosphate buffered saline (PBS), cells were incubated with an anti-human IgG-FITC conjugate (Sigma) for 1 h at 4°C. After washing, propidium iodide (PI) was added to cells (Sigma), with PI-negative.
Downstream tips
Cells were analyzed for bound antibody using a FACSCalibur flow cytometer (Becton-Dickinson, USA). Data acquisition and analyses were with the CellQuest software (Becton-Dickinson).

Publication protocol

For analysis of L1CAM expression at the cell surface, cells were incubated with the human anti-L1CAM mAb, Ab417 (Cho et al., 2016), for 1 h at 4°C. After washing twice with phosphate buffered saline (PBS), cells were incubated with an anti-human IgG-FITC conjugate (Sigma) for 1 h at 4°C. After washing, propidium iodide (PI) was added to cells (Sigma), with PI-negative cells subsequently analyzed for bound antibody using a FACSCalibur flow cytometer (Becton-Dickinson, USA). Data acquisition and analyses were with the CellQuest software (Becton-Dickinson).

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