Bolttm 4 12 bis tris gels

Full length antibody, purified F(ab)'2 and purified Fc fragments were diluted in NuPAGE LDS sample buffer (#NP0007, Invitrogen), heated at 70 °C for 10 min and loaded onto Bolt^TM 4-12% Bis-Tris gels (#NW04120BOX, Invitrogen). Electrophoresis was run on the Mini Gel Tank system (Life Technologies) at 200V constant voltage in NuPAGE MES SDS running buffer (#NP0002, Invitrogen). Gels were fixed and stained with Coomassie brilliant blue R-250 (#1610436, Bio-Rad).

Full-length monoclonal CD4 antibody (IgG), reduced full-length CD4 antibody and the tracer [⁶⁴Cu]Cu-NOTA-CD4 were mixed with NuPAGE LDS sample buffer (#NP0007, Invitrogen, Carlsbad, CA, USA) and denatured for 10 min at 70 °C and loaded onto BoltTM 4–12% Bis–Tris gels (#NW04120, Invitrogen). Electrophoresis was run on the Mini Gel Tank (Thermo Fisher Scientific, Waltham, MA, USA) at 200 V constant voltage in NuPAGE MES SDS running buffer (#NP0002, Invitrogen). The SDS-PAGE gel was fixed and stained using Coomassie brilliant blue R-250 (#1610436, Bio-Rad, Hercules, CA, USA). The gel was analyzed for radioactive content by exposure to Multisensitive Phosphor Screens and imaged using the Amersham Typhoon Imaging System (GE Healthcare, Chicago, IL, USA).