Porcine peripheral red blood cells were paraffn embedded and sectioned. 4 um sections were de-paraffnized and rehydrated. Sections were incubated with or without additional thiol compounds and L-Cysteine monoclonal antibody, clone F2D (Cat # MAB6677) for 2 hours at room temperature. Antibody concentration was 3 ug/mL . Sections were blocked pre- and post antibody treatements with 1% BSA in PBS. Development was with a goat anti-mouse IgG antibody labeled with Alexa™ 488.
Cysteine addition was carried out at 20 mM in 1% BSA in PBS and added with L-Cysteine monoclonal antibody, clone F2D (Cat # MAB6677) at 3 ug/mL for 1 hour at room temperature. With polymorphonuclear neutrophils addition of 20 mM cytseine reduced or eliminated the goat anti-mouse IgG-Alexa 488 signal Nuclear staining was carried out with TOPRO at 1 : 500 dilution in PBS.