Anti vsv g clone p5d4

Anti‐PlexinB2 antibody used for Western blotting analysis was purchased from Abcam (ab193355; dilution 1:500). EGFR antibody was provided by Enzo Life Sciences (ALX‐804‐064‐C100; dil. 1:500), while anti‐phospho‐EGFR‐specific antibodies (directed to p‐Tyr1068; ab5644; dil. 1:500) were from Abcam. Anti‐vinculin (V4505; dil. 1:1,000) and anti‐VSV‐G (clone P5D4; dil. 1:1,000) were provided by Sigma‐Aldrich. Anti‐phospho‐p44/42‐MAPK (Erk1/2; Thr202/Tyr204; #4370; dil. 1:1,000), anti‐p44/42‐MAPK (Erk1/2; 137F5; #4695; dil. 1:1,000), and anti‐phospho‐S6 ribosomal protein (Ser240/244; #2215; dil. 1:1,000) antibodies were from Cell Signaling.

Cell lysates from P. gingivalis WT or mutants were separated by SDS-PAGE, and the proteins were transferred onto a nitrocellulose membrane. The membrane was blocked in 5% skim milk and probed with MAb-1B5 (a kind gift from M.A. Curtis) (51 (link)), Sov-, PPAD- or CPG70-specific antibodies, or commercial anti-FLAG (clone M2, Sigma-Aldrich), and anti-VSV-G (clone P5D4, Sigma-Aldrich) monoclonal antibodies followed by anti-mouse or anti-rabbit HRP (or alkaline phosphatase)-conjugated secondary antibodies. HRP was developed using SuperSignal West Pico chemiluminescent substrate and visualized with an LAS-3000 imaging system. Alkaline phosphatase signal was developed in alkaline buffer in the presence of 5-bromo-4-chloro-3-indolylphosphate and nitroblue tetrazolium.