cDNAs for the anti-AGIA antibody heavy and light chains [17 (link)] were subcloned into the pcDNA3.4 expression vector using PCR and In-Fusion Reaction. Anti-AGIA antibody was expressed using the Expi293F Expression System (Thermo Fisher Scientific) according to the manufacturer’s instruction. The antibody secreted in culture medium was purified by protein G sepharose 4 Fast Flow (GE Healthcare), and then buffer exchange was conducted using a PD-10 column. Purified antibody was frozen and stored at -20°C.
Preparations of antibody conjugated sepharose beads was conducted using NHS-activated Sepharose 4FF (GE Healthcare). Anti-AGIA antibody sepharose was stored at 4°C. HRP-conjugated antibody was prepared using the HRP Conjugation Kit (Abcam). Anti-AGIA antibody-HRP was stored at -30°C.
Preparations of antibody conjugated sepharose beads was conducted using NHS-activated Sepharose 4FF (GE Healthcare). Anti-AGIA antibody sepharose was stored at 4°C. HRP-conjugated antibody was prepared using the HRP Conjugation Kit (Abcam). Anti-AGIA antibody-HRP was stored at -30°C.
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