IgG (Fab region), Jackson ImmunoReseach Europe, 115-605-006

I have used this abtibody to stain T cells for the expression of a chimeric receptor (CAR T cells). Cells were transduced with a lentiviral vector containing a GFP reporter gene.

Protocol:
Around 300.000 cells were stained with dead stain (THERMO FISHER SCIENTIFIC, L10119) and 6 uL of F(ab’)2 specific antibody diluted 1:10 in PBS + 2 % FBS. Cells were stained for 30 minutes at 4 degress in the dark.
Cells were then washed four times with 2 mL PBS + 2 % FBS before being stained with surface marker antibodies CD3 (BD, 563800), CD4 (BD, 562970) and CD8 (BD, 566860) (this step is important in order to avoid unwanted binding to mouse anti-human antibodies). Cells were washed once before running on flow.

Antibody has been titrated and compensation was made for flow cytometry using cells and an anti-human HLA-A, B, C antibody (Biolegend, 311412).