α rat igg

As primary antibodies we used: mouse monoclonal α-histidine (α-His; Roche), mouse monoclonal α-BtUb (α-Ub; SantaCruz), rabbit α-EhADH (α-EhADH18) (Díaz-Hernández et al., 2021 (link)), mouse α-EhVps32 (α-EhVps32) (Avalos-Padilla et al., 2015 (link)), mouse α-LBPA (α-LBPA; Echelon Bioscience) and mouse monoclonal α-human actin (α-actin) (kindly given by Dr. Manuel Hernández, Cell Biology Department, CINVESTAV IPN). Secondary antibodies were: HRP-labelled α-rabbit, mouse and rat IgGs (Zymed) for western blot. FITC-labelled α-rabbit and α-mouse IgGs and TRITC-labelled α-rat IgGs (Life Technologies) for immunofluorescence. For immunoelectron microscopy experiments, we used α-rat IgG, conjugated with 10 nm gold particles and α-mouse IgG, conjugated with 20 nm gold particles (TED Pella Inc).

For EhADH immunodetection, we obtained hamster polyclonal antibodies (α-EhADH) against a specific EhADH peptide (N₅₆₆-QCVINLLKEFDNTKNI-C₅₈₂) localized within the adherence domain. Male hamsters were immunized three times (each 2 weeks) with 300 μg of this peptide diluted in TiterMax^® Gold Adjuvant liquid (Sigma), then, animals were bled and antibodies obtained. Other primary antibodies used were: rat α-EhVps23 (Galindo et al., 2021 (link)), mouse monoclonal α-HA (GeneTex), mouse α-EhVps32 (Avalos-Padilla et al., 2015 (link)), mouse monoclonal α-Ubiquitin (Santacruz), rabbit α-EhCP112 (García-Rivera et al., 1999 (link)) and mouse monoclonal α-human actin (kindly given by Dr. Manuel Hernández, Cell Biology Department, CINVESTAV) antibodies. Secondary antibodies were: HRP-labelled α-rabbit, α-mouse, α-hamster and α-rat IgGs (Zymed) for western blot. FITC-labelled α-rabbit and α-mouse IgGs, Alexa 647 α-hamster and TRITC-labelled or Alexa 405 α-rat IgGs (Life Technologies) for immunofluorescence. For immunoelectron microscopy experiments, we used α-rat IgG, conjugated with 10 nm gold particles (TED Pella Inc).