Western blot was performed using Flotillin 2 Polyclonal Antibody (Product # PA5-17178) and a 53 kDa band corresponding to FLOT2 was observed in HeLa, A-431, NIH/3T3 and Mouse Brain along with an uncharacterized band (*) at ~80 kDa in A-431. Whole cell extracts (30 µg lysate) of HeLa (Lane 1), A-431 (Lane 2), Molt-4 (Lane 3), NIH/3T3 (Lane 4) and tissue extracts (30 µg lysate) of Mouse Brain (Lane 5) were electrophoresed using NuPAGE™ 12% Bis-Tris Protein Gel (Product # NP0341BOX) 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 (1:1000 dilution) and detected by chemiluminescence with Goat anti-Rabbit IgG (Heavy Chain), Superclonal™ Recombinant Secondary Antibody, HRP (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).