Overlay histogram showing SK-OV-3 cells stained with BD-PB1695(green line). The cells were fixed with 2% paraformaldehyde and then permeabilized with 90% methanol for 10 min. The cells were then icubated in 2% bovine serum albumin to block non-specific protein-protein interactions followed by the antibody (1:25 dilution) for 60 min at 37ºC. The secondary antibody used was Goat-Anti-Rabbit IgG, DyLight® 488 Conjugated Highly Cross-Adsorbed at 1/200 dilution for 40 min at Room temperature. Isotype control antibody (blue line) was rabbit IgG1 (1μg/1x10^6 cells) used under the same conditions. Acquisition of >10, 000 events was performed.

Anti-SPNS2 Antibody (N-term) at 1:1000 dilution + SK-OV-3 whole cell lysate Lysates/proteins at 20 µg per lane. Secondary Goat Anti-Rabbit IgG, (H+L), Peroxidase conjugated at 1/10000 dilution. Predicted band size : 58 kDa Blocking/Dilution buffer: 5% NFDM/TBST.

All lanes : Anti-SPNS2 Antibody (N-term) at 1:1000 dilution Lane 1: Human heart lysate Lane 2: Human kidney lysate Lane 3: Human skeletal muscle lysate Lane 4: SK-BR-3 whole cell lysate Lysates/proteins at 20 µg per lane. Secondary Goat Anti-Rabbit IgG, (H+L), Peroxidase conjugated at 1/10000 dilution. Predicted band size : 58 kDa Blocking/Dilution buffer: 5% NFDM/TBST.