Fig. 1: Biotinylation of AGIA-tag protein by AGIA-FabID and AGIA-mAbID. | Nature Communications

Fig. 1: Biotinylation of AGIA-tag protein by AGIA-FabID and AGIA-mAbID.

From: Proximity extracellular protein-protein interaction analysis of EGFR using AirID-conjugated fragment of antigen binding

Fig. 1

a Schematic of AGIA antibody fused with the proximity biotinylation enzyme AirID. AirID is fused to AGIA-Fab or AGIA-mAb, b SDS-PAGE and Coomassie Brilliant Blue (CBB) staining in reducing and non-reducing state of AGIA-FabID and AGIA-mAbID synthesised by Expi293F expression system and purified by Ni Sepharose excel. c Detection of self-biotinylation of AGIA-FabID and AGIA-mAbID by immunoblotting using an anti-biotin antibody. Detection of AGIA-FabID and AGIA-mAbID using an anti-His tag antibody. d Schematic of biotinylation targeting the AGIA-tagged protein using AGIA-FabID and AGIA-mAbID. e Comparison of the activities of AGIA-FabID and AGIA-mAbID targeting FLAG-GST-DRD1 CTD (FLAG-GST fused to the C-terminus of DRD1) in vitro. f Targeting various AGIA-tagged proteins biotinylated by AGIA-FabID. g Schematic of the biotinylation-targeting complex of p53-MDM2 using AGIA-FabID. h Confirmation of biotinylation of FLAG-GST-MDM2 and AGIA-p53 by immunoblotting blotting. The SHB and LHB are S-protein of hepatitis B virus. b, c, e, f, h, The experiment was independently repeated twice, with similar results. Source data are provided as a Source Data file.

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