Lewis rat splenocytes stained with ab6413 (right) or mouse IgG1κ (ab170190) isotype (left). Lewis rat splenocytes were incubated for 30 min on ice in 1x PBS / 10 % rat serum to block FC receptors and non-specific protein-protein interaction followed by the antibody (ab6413) or mouse IgG1κ (ab170190) (1x¹⁰⁶ in 100 µl at 0.2 µg/ml) for 30 min on ice.

The secondary antibody Goat anti-mouse IgG H&L (Alexa Fluor ^® 488, pre-adsorbed) (ab150117) was used at 1/2000 dilution for 30 min on ice. The cells were simultaneously stained with CD3 antibody.

Acquisition of >30,000 events were collected using a 50 mW Blue laser (488nm) and 530/30 bandpass filter. Events were gated on viable lymphocytes.