Protein extraction
was carried out from P. gingivalis cells (control
and treated) as previously reported.19 (link) Briefly,
the cells were centrifuged for 5 min at 5,000g at
4 °C. Then, the supernatants were discarded, and the resulting
pellets were washed with phosphate-buffered saline (PBS). Subsequently,
the protein pellets were suspended in lysis buffer (0.5 mL; pH 8.8;
30 mM Tris buffer containing 7 M urea, 2 M thiourea, 2% Chaps, and
the protease inhibitor cocktail; GE Healthcare, Chicago, IL, USA)
on ice for 20 min. After that, samples were vortexed and sonicated
(3–4 times) for 1 min and recentrifuged at 10,000g at 4 °C for 3 min to remove cell debris. Finally, protein concentrations
were determined using the 2D-Quant Kit according to the manufacturer’s
instructions (GE Healthcare).
was carried out from P. gingivalis cells (control
and treated) as previously reported.19 (link) Briefly,
the cells were centrifuged for 5 min at 5,000g at
4 °C. Then, the supernatants were discarded, and the resulting
pellets were washed with phosphate-buffered saline (PBS). Subsequently,
the protein pellets were suspended in lysis buffer (0.5 mL; pH 8.8;
30 mM Tris buffer containing 7 M urea, 2 M thiourea, 2% Chaps, and
the protease inhibitor cocktail; GE Healthcare, Chicago, IL, USA)
on ice for 20 min. After that, samples were vortexed and sonicated
(3–4 times) for 1 min and recentrifuged at 10,000g at 4 °C for 3 min to remove cell debris. Finally, protein concentrations
were determined using the 2D-Quant Kit according to the manufacturer’s
instructions (GE Healthcare).
Free full text: Click here
