Cellular Fractionation of H. pylori

Cellular fractionation of the wild‐type H. pylori cells was performed as described earlier 14 (link). Briefly, bacterial cell extract was centrifuged at 148 000 g in SW‐55 rotor (Beckman coulter ultracentrifuge, Irving, TX, USA) at 4 °C for 1 h, the supernatant was recovered as cytoplasmic/periplasmic fraction (C/P) and the resulting pellet was recovered as total membrane (TM). Volume of C/P was measured and TM was resuspended in the same volume of PBS. Equal volume of each sample was mixed with 2X‐SDS sample buffer, separated in SDS/PAGE, and subjected to western blotting using an appropriate antibody.