Anti lptd

For immunoblot analysis, total membrane proteins (25 μg) resolved by SDS-PAGE under nonreducing (in the absence of β-mercaptoethanol) conditions (37 (link)) as described above were electrotransferred onto an Immun-Blot polyvinylidene difluoride (PVDF) membrane (Bio-Rad) and probed for LptD or LptE using anti-LptD (1:20,000) and anti-LptE (1:30,000) polyclonal rabbit antibodies (GenScript) and goat anti-rabbit IgG horseradish peroxidase (HRP)-conjugated secondary antibody (Sigma-Aldrich). Anti-LptD and anti-LptE antibodies were generated at GenScript using keyhole limpet hemocyanin (KLH)-conjugated peptides as the antigens (LptD epitope, QLHQKEAPGQPEPVC, and LptE epitope, ELLDKETTRKDVPSC). The epitopes were selected using the OptimumAntigen design tool (GenScript). The transferred membrane was developed with Clarity Western ECL substrate (Bio-Rad) and visualized in a FluorChem Q imager (ProteinSimple).

For immunoblot analysis, total membrane proteins (25 μg) resolved by SDS-PAGE under nonreducing (in the absence of β-mercaptoethanol) conditions (37 (link)) as described above were electrotransferred onto an Immun-Blot polyvinylidene difluoride (PVDF) membrane (Bio-Rad) and probed for LptD or LptE using anti-LptD (1:20,000) and anti-LptE (1:30,000) polyclonal rabbit antibodies (GenScript) and goat anti-rabbit IgG horseradish peroxidase (HRP)-conjugated secondary antibody (Sigma-Aldrich). Anti-LptD and anti-LptE antibodies were generated at GenScript using keyhole limpet hemocyanin (KLH)-conjugated peptides as the antigens (LptD epitope, QLHQKEAPGQPEPVC, and LptE epitope, ELLDKETTRKDVPSC). The epitopes were selected using the OptimumAntigen design tool (GenScript). The transferred membrane was developed with Clarity Western ECL substrate (Bio-Rad) and visualized in a FluorChem Q imager (ProteinSimple).