Extended Data Fig. 4. Light dependent covalent bond formation between 7D12 and EGFR is antibody-specific.
For these experiments, the amount of EGFR is 10 picomoles, incubation time prior to irradiation is 5 min, irradiation time with 365 nm light is 10 min, and the amount of 7D12 and its Bpa-containing mutants is 100 picomoles. (A) In the Coomassie stained gel image, the band higher than EGFR is photocrosslinked 7D12-109Bpa + EGFR complex. This is the full gel image for data shown in Fig. 2e. Lane marked L is the Invitrogen SeeBlue Plus2 Pre-stained Protein Standard (Catalog no. LC5925). These results demonstrate that the photocrosslinking occurs only for 7D12-109Bpa upon irradiation with 365 nm light. (B) Percentage crosslinked product calculated by measuring the intensity of the band corresponding to 7D12-109Bpa + EGFR and EGFR. These experiments were repeated twice with similar results.