Photo-conjugation efficiencies for the different Z domain probes to mouse IgG1.

The efficiencies of photoconjugation were analysed through scanning of the gel images in Figure 5 using ImageJ software (see Materials and methods section). (A) Photo-conjugation efficiencies for 16 probe variants, derived from either the ZWT-domain (white bars) or the ZF5I-domain, containing the F5I substitution (black bars). The position at which a cysteine was introduced to site-specifically position the MBP probe is indicated for each probe. (B) Mean values of photo-conjugation efficiencies (%) from the use of either the ZWT-Q32C-MBP (white bar; mean value = 45.4±4.1%) or ZF5I-Q32C-MBP (black bar; mean value = 64.4±5.3%) probes to a large panel of different mIgG1 antibodies (16 of the mAbs seen in Figure 5B).