Western Blot analysis of IKAROS was performed by separating 30 µg of various whole cell extracts by 7.5% SDS-PAGE. Proteins were transferred to a membrane and probed with a IKAROS Polyclonal Antibody (Product # PA5-85570) at a dilution of 1:500 and a HRP-conjugated anti-rabbit IgG secondary antibody.

Western blot was performed using Anti-IKAROS Polyclonal Antibody (Product # PA5-85570) and 60, 52 and 40 k Da bands corresponding to IKZF1 along with an uncharacterized band (*) was observed across positive cell lines (REH, Daudi, Ramos and Jurkat). Whole cell extracts (30 µg lysate) of REH (Lane 1), Daudi (Lane 2), Ramos (Lane 3), Jurkat (Lane 4), U-937 (Lane 5), HL-60 (Lane 6), SH-SY5Y (Lane 7) and A-431 (Lane 8) were electrophoresed using NuPAGE™ 10% Bis-Tris Protein Gel (Product # NP0301BOX). 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 # A27036,1:20000 dilution) using the iBright™ FL1500 Imaging System (Product # A44115). Chemiluminescentdetection was performed using Novex® ECL Chemiluminescent Substrate Reagent Kit (Product # WP20005).