Immunohistochemical analysis of Ferritin Light Chain in paraffin-embedded Ca922 xenograft using a Ferritin Light Chain polyclonal antibody (Product # PA5-29599) at a dilution of 1:100.

Western blot was performed using Anti-Ferritin Light Chain Polyclonal Antibody (Product # PA5-29599) and a 20kDa band corresponding to Ferritin Light Chain was observed across tissues tested with lower levels in mouse spleen. Tissue extracts (30 µg lysate) of Mouse Liver (Lane 1), Rat Liver (Lane 2), Mouse Spleen (Lane 3) 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). Ferritin is a holoenzyme shell with a molecular weight of about 450 kDa, and consists of 24 subunits of two types, H(heavy) and L(light), hence band above 260kDa could correspond to the holoenzyme shell of 450kDa.