Western blot was performed using Anti-S100A6 Polyclonal Antibody (Product # PA5-80990) and a 10kDa band corresponding to Protein S100-A6 was observed. Whole cell extracts (30 µg lysate) of HeLa (Lane 1), A549 (Lane 2), PC-3 (Lane 3), U-87 MG (Lane 4), HEK-293 (Lane 5), K-562 (Lane 6) and LNCaP (Lane 7) were electrophoresed using Novex™ 16% Tricine Protein Gel (Product # EC6695BOX). 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) and detected by chemiluminescence with Goat anti-Rabbit IgG (Heavy Chain) Superclonal™ Recombinant Secondary Antibody, HRP (Product # A27036, 1:4000) using the iBright FL 1000 (Product # A32752). Chemiluminescent detection was performed using Novex® ECL Chemiluminescent Substrate Reagent Kit (Product # WP20005). Band of interest was not observed in HEK-293, K-562 and LNCaP which are reported to be either negative or express S100A6 at very low levels.