Primary antibodies were: (1) FXYD12: a rabbit polyclonal antibody (LTK BioLaboratories, Taoyuan, Taiwan) targeting the C-terminus of medaka FXYD12 (S2 Fig ); (2) NKA: a mouse monoclonal antibody (α5; Developmental Studies Hybridoma Bank, Iowa City, IA, USA) against the α-subunit of avian NKA; and (3) actin: a rabbit polyclonal antibody (sc-1616-R; Santa Cruz Biotechnology, Santa Cruz, CA, USA) against the C-terminus of human actin, as a loading control for immunoblotting. The specificity and related information for these antibodies have been demonstrated and described previously [25 (link), 29 ].
The secondary antibodies employed for immunofluorescent staining were Alexa-Fluor-546-conjugated goat anti-rabbit IgG and Alexa-Fluor-488-conjugated goat anti-mouse IgG (Molecular Probes, Eugene, OR, USA). For immunoblotting, the secondary antibodies were horseradish-peroxidase-conjugated goat anti-mouse IgG and anti-rabbit IgG (Pierce, Rockford, IL, USA).
The secondary antibodies employed for immunofluorescent staining were Alexa-Fluor-546-conjugated goat anti-rabbit IgG and Alexa-Fluor-488-conjugated goat anti-mouse IgG (Molecular Probes, Eugene, OR, USA). For immunoblotting, the secondary antibodies were horseradish-peroxidase-conjugated goat anti-mouse IgG and anti-rabbit IgG (Pierce, Rockford, IL, USA).
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