Western blot was performed using Anti-NCKAP1 Rabbit Polyclonal Antibody (Product # PA5-21680) and a 130kDa band corresponding to NCKAP1 was observed across cell lines and tissues tested except U-937 and HL-60 which are reported negative for NCKP1 expression. An uncharacterized band was also observed at ~30kDa in certain cell lines. Membrane enriched extracts (30 µg lysate) of U-87 MG (Lane 1), NTERA-2 cl.D1 (Lane 2), A-431 (Lane 3), U-2 OS (Lane 4), Mouse Brain (Lane 5), Rat Brain (Lane 6), U-937 (Lane 7) and HL-60 (Lane 8) were electrophoresed using Novex® NuPAGE® 4-12 % Bis-Tris gel (Product # NP0322BOX). Resolved proteins were then transferred onto a nitrocellulose membrane (Product # IB23001) by iBlot® 2 Dry Blotting System (Product # IB21001). The blot was probed with the primary antibody (1:2000 dilution) and detected by chemiluminescence Goat Anti-Rabbit IgG Secondary Antibody, HRP conjugate (Product # A27036, 1:4000 dilution) using the iBright FL 1000 (Product # A32752). Chemiluminescent detection was performed using Novex® ECL Chemiluminescent Substrate Reagent Kit (Product # WP20005).