Western blot was performed using Anti-CD64 Recombinant Rabbit Monoclonal Antibody (27) (Product # MA5-29706) and a 65kDa band corresponding to CD64 was observed. Membrane enriched extracts (30 µg lysate) of RAW 264.7 (Lane 1), NIH/3T3 (Lane 2) and Neuro-2a (Lane 3) were electrophoresed using NuPAGE™ 4-12% Bis-Tris Protein 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 (1:1000) 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 is observed only in RAW 264.7, a macrophage cell line that expresses CD64 constitutively.