Fig. 1. Characterisation of the complex between antibody 7D10 and both BAK and BAK^L100A.

a Size exclusion chromatography demonstrates that the 7D10 Fv fragment forms a 1:1 complex with wild-type BAK and with BAK^L100A variant. (BAK is ΔN22ΔC25Δcys 6X His, i.e. with N- and C-terminal truncations, with wild-type cysteine residues replaced with serine and a C-terminal His-tag). BSA, bovine serum albumin, MW 66 kDa. b, c Both the 7D10 antibody and its Fv fragment can permeabilise liposomes to which BAK or BAK L100A has been immobilised via a C-terminal His-tag. Results show the mean (line) ± SEM (shaded area) for three independent experiments with at least two technical replicates. BAK is used at 150 nM. Controls shown for activation by BIM BH3 peptide reproduce previous results [16].