Figure 7
From: Assembly and Folding Properties of Cytosolic IgG Intrabodies
IgG1s expressed in the reducing cytosol and via the oxidizing ER assemble differently. (a) The concentration of free thiols in PBS was measured before and after DTT removal steps. (b) Immunoblotting analysis of the assembly pattern of H and L chains following DTT removal. HEK293 cells transfected with KV10ΔLd-3D8 IgG or KV10Ld-3D8 IgG. Lysates of transfectants were treated with 100 mM DTT for 30 min at room temperature, followed by DTT removal steps comprising passage through a desalting column and dialysis against PBS at 4 °C for 24 hours. Lysates were separated by non-reducing SDS-PAGE, and the H chain was detected by immunoblotting with anti-IgG/Fc antibody.
