Rabbit anti prrt2

These experiments were performed as described previously (27 (link)). Primary antibodies were as follows: rabbit anti-PRRT2 (1:500, Sigma-Aldrich), mouse anti-actin (1:1000, Sigma-Aldrich), mouse anti-Na⁺/K⁺ATPase (1:1000, Millipore), and mouse anti-dynamin I (1:1000, Millipore).

Cells transfected with the various PRRT2 constructs were live-labeled by diluting primary antibodies (rabbit anti-PRRT2, 1:200, Sigma-Aldrich; mouse anti-HA, 1:200, Invitrogen; mouse anti-turboGFP, 1:200, Origene; and mouse anti-GFP, 1:500, Millipore) in culture medium for 1 h at 37 °C in a 5% CO₂ incubator to detect the surface epitopes, followed by incubation with Alexa Fluor 488 or 594 secondary antibodies for 1 h at 37 °C, and fixed. In the experiments of live-labeling with HA antibodies, after live incubation with Alexa Fluor 488 secondary antibodies, cells were fixed with 4% paraformaldehyde, permeabilized with 0.1% Triton X-100, and stained with anti-PRRT2 antibodies, followed by Alexa Fluor 594 secondary antibodies to label total PRRT2.