Western blot analysis was performed on nuclear enriched extracts of A549 (Lane 1), THP-1 (Lane 2), Jurkat (Lane 3), Ramos (Lane 4), HEK-293 (Lane 5) and tissue extract of Rat Testis (Lane 6). The blot was probed with Anti-RANBP3 Rabbit Polyclonal Antibody (Product # PA1-084, 2µg/mL) and detected by chemiluminescence using Goat anti-Rabbit IgG (Heavy Chain) Superclonal™ Secondary Antibody, HRP conjugate (Product # A27036, 0.25µg/mL, 1:4000 dilution). A 60 kDa band corresponding to RANBP3 was observed across cell lines and tissue tested. Known quantity of protein samples were electrophoresed using Novex® NuPAGE® 4-12 % Bis-Tris gel (Product # NP0321BOX), XCell SureLock™ Electrophoresis System (Product # EI0002) and Novex® Sharp Pre-Stained Protein Standard (Product # LC5800). Resolved proteins were then transferred onto a nitrocellulose membrane with iBlot® 2 Dry Blotting System (Product # IB21001). The membrane was probed with the relevant primary and secondary Antibody following blocking with 5 % skimmed milk. Chemiluminescent detection was performed using Pierce™ ECL Western Blotting Substrate (Product # 32106).

Immunohistochemical staining of RanBP3 in NRK cells using Product # PA1-084.