Figure 3
C1‐INH conformers in reducing and non‐reducing SDS-PAGE. (a) Plasma samples (NHP and 378 R/C) and recombinant C1-INH expressed in COS7 cells (rwt for the wild‐type construct and r378C for the mutant) were electrophoresed in reducing (+DTT, right) and not reducing (−DTT, left) SDS‐PAGE before blotting. When the disulphide bonds were reduced the higher bands disappeared. Normal human plasma (NHP) and kaolin activate plasma (NHP*) are shown as reference. (b) Recombinant C1‐INH present in cell media (S: secreted) or in the cell post‐nuclear supernatant (I: intracellular) were electrophoresed in reducing (right) and not reducing (left) SDS‐PAGE before blotting. Bands corresponding to monomeric (105 kd for the secreted form, slightly less for the intracellular, not fully glycosylated, form) and dimeric (around 250 kd) forms were indicated with arrowheads and arrows respectively. The primary Ab used was a commercial polyclonal antibody against C1‐INH (The Binding Site Ltd, Birmingham, UK).
