*4.7. Whole Cell Lysates*

Harvested cells were placed on ice. Three hundred to four hundred microliters of ice-cold lysis buffer (0.15-M NaCl, 0.05-M Tris, pH 8.0, 1% (*v*/*v*) Triton-X100, 1-mM EDTA, and 1-mM pepstatin, 1.3-mM benzamidine, and 1.8-mM leupeptin) was added to the cells. The cells were resuspended by vortexing. The cell suspension obtained was kept on ice and vortexed every 10 min for 1 h. Cells suspension were then centrifuged for 15 min at 15,600× *g* at 4 ◦C, and the supernatant containing the whole cell lysate was quantified for total protein concentrations using a bicinchoninic acid kit (Pierce, ThermoFisher Scientific). The samples were mixed with sample buffer for loading on sodium dodecyl sulphate (SDS) gels.
