Western blot was performed using F(ab)2-Goat anti-Human IgG, IgM, IgA (H+L) Secondary Antibody (Product # A24496). 55-80 kDa and 25 kDa bands corresponding to Human Ig heavy and light chains respectively were observed for IgG1, IgG2, IgG3, IgG4, IgA, IgM and Mouse IgM. No reactivity was observed for Human IgE heavy chain, Mouse IgG and Rabbit IgG (Panel I). Purified protein (100 ng) of Human IgG1 (Lane 1), IgG2 (Lane 2), IgG3 (Lane 3), IgG4 (Lane 4), IgA (Lane 5), IgM (Lane 6), IgE (Lane 7), Mouse IgG (Lane 8), Mouse IgM (Lane 9) and Rabbit IgG (Lane 10) were electrophoresed using NuPAGE™ 4-12% Bis-Tris Protein Gel (Product # NP0322BOX). Resolved proteins were then transferred to a nitrocellulose membrane (Product # IB23001) by iBlot® 2 Dry Blotting System (Product # IB21001). The blot was probed with F(ab)2-Goat anti-Human IgG, IgM, IgA (H+L) Secondary Antibody (Product # A24496) (1:2000 dilution) and detected by chemiluminescence with Rabbit anti-Goat IgG F(ab)2 Secondary Antibody, HRP (Product # 31403, 1:20000 dilution) using iBright FL1500 (Product # A44115). Chemiluminescentdetection was performed using Novex® ECL Chemiluminescent Substrate Reagent Kit (Product # WP20005). Silver staining was performed to establish equivalent loading using Pierce™ Silver Stain Kit (Product # 24612) (Panel II). BSA (*), part of Mouse IgM formulation, can be seen at ~60 kDa.

Western blot was performed using F(ab)2-Goat anti-Human IgG, IgM, IgA (H+L) Secondary Antibody (Product # A24496). 55-80 kDa and 25 kDa bands corresponding to Human Ig heavy and light chains respectively were observed for IgG1, IgG2, IgG3, IgG4, IgA, IgM and Mouse IgM. No reactivity was observed for Human IgE heavy chain, Mouse IgG and Rabbit IgG (Panel I). Purified protein (100 ng) of Human IgG1 (Lane 1), IgG2 (Lane 2), IgG3 (Lane 3), IgG4 (Lane 4), IgA (Lane 5), IgM (Lane 6), IgE (Lane 7), Mouse IgG (Lane 8), Mouse IgM (Lane 9) and Rabbit IgG (Lane 10) were electrophoresed using NuPAGE™ 4-12% Bis-Tris Protein Gel (Product # NP0322BOX). Resolved proteins were then transferred to a nitrocellulose membrane (Product # IB23001) by iBlot® 2 Dry Blotting System (Product # IB21001). The blot was probed with F(ab)2-Goat anti-Human IgG, IgM, IgA (H+L) Secondary Antibody (Product # A24496) (1:2000 dilution) and detected by chemiluminescence with Rabbit anti-Goat IgG F(ab)2 Secondary Antibody, HRP (Product # 31403, 1:20000 dilution) using iBright FL1500 (Product # A44115). Chemiluminescentdetection was performed using Novex® ECL Chemiluminescent Substrate Reagent Kit (Product # WP20005). Silver staining was performed to establish equivalent loading using Pierce™ Silver Stain Kit (Product # 24612) (Panel II). BSA (*), part of Mouse IgM formulation, can be seen at ~60 kDa.