Western blot was performed using AHNAK Monoclonal Antibody (EM-09) (Product # MA1-10050) and a 680 kDa band corresponding to AHNAK was observed across cell lines tested. Whole cell extracts (60 µg lysate), prepared with RIPA buffer, of U-87 MG (Lane 1), PANC-1 (Lane 2), 769-P (Lane 3), NIH:OVCAR-3 (Lane 4), HaCaT (Lane 5), SW480 (Lane 6), Jurkat (Lane 7), A-673 (Lane 8), HEL 92.1.7 (Lane 9) were electrophoresed using NuPAGE™ 4-12% Bis-Tris Protein Gel (Product # NP0321BOX), 10 well. Resolved proteins were then transferred onto a nitrocellulose membrane (Product # IB23002) by iBlot® 2 Dry Blotting System (Product # IB21001). The blot was probed with the primary antibody (1:2000) and detected by chemiluminescence with Goat anti-Mouse IgG (H+L) Superclonal™ Recombinant Secondary Antibody, HRP (Product # A28177, 1:20,000) using the iBright™ FL1500 Imaging System (Product # A44115). Chemiluminescent detection was performed using SuperSignal™ West Atto Ultimate Sensitivity Substrate (Product # A38556). As reported, the levels of AHNAK were low in Jurkat, A-673 and HEL.92.17 cell lines as shown here in the figure.