2.5. Sensitization of latex bead with GST-LipL32 recombinant proteins

Latex polystyrene beads, 0.8 μM diameter (Sigma, USA) were sensitized with recombinant GST-LipL32 protein according to standard method. Briefly, latex bead suspension was washed twice with carbonate-bicarbonate buffer. After that, latex suspension was incubated with 400 μg of GST-LipL32 recombinant protein at 37 °C for 6 h. The sensitized beads were washed and suspended in PBS containing 5 mg/mL of BSA to final concentration as 2%. The latex mixers were left overnight at 37 °C. Finally, latex suspension was washed again and suspended with PBS containing 0.5 mg/mL of BSA and 0.1% sodium azide to a 1% latex suspension. The sensitized latex beads were stored at 4 °C until examination.