Immunohistochemistry and Protein Interaction Assays

Immunohistochemistry: anti-RBP2 (guinea pig polyclonal, 1:1000; kind gift of Eckart Gundelfinger; specifically recognizes RBP2 as verified by the knockout, [21 (link)]); anti-Cav1.3 (rabbit polyclonal, 1:500; Alomone Labs; [53 (link)]); anti-CtBP2/RIBEYE (mouse monoclonal, 1:200; BD Biosciences); Alexa488 anti-guinea pig (goat polyclonal; 1:500; Invitrogen), Alexa568 anti-mouse (goat polyclonal; 1.500; Invitrogen), Cy3 anti-rabbit (donkey polyclonal; 1:1500; Jackson Immuno Research); co-immunoprecipitation: affinity purified anti-RBP2-1318 (rabbit polyclonal; 1:100; directed against amino acids 261–284, NCBI reference sequence NP_001074857.1); anti-GFP (mouse monoclonal; 4 μl of 0.4 μg/μl; Roche); peroxidase-conjugated secondary antibodies: anti-rabbit (goat polyclonal; 1:20,000; Sigma-Aldrich); anti-mouse (goat polyclonal; 1:5000; Sigma-Aldrich); GST (glutathione-S-transferase) pull-down: anti-GAPDH (rabbit monoclonal; 1:1000; Cell Signaling Technology); anti-HA (mouse monoclonal κ16B12; 1:1000; Biozym); affinity-purified anti-Cav1.3α1_2022–2138 (rabbit polyclonal; 1:1000; [51 (link)]). Peroxidase-conjugated secondary antibodies: anti-mouse (polyclonal goat; 1:3000; Roth) and anti-rabbit (polyclonal goat; 1:3000; Roth).