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. 2021 Dec 6;12:742875. doi: 10.3389/fpls.2021.742875

FIGURE 2.

FIGURE 2

Sodium dodecyl-sulfate polyacrylamide gel electrophoresis (SDS-PAGE) (A) and Western blot (B,C) analysis of plant produced, Ni-NTA resin purified gACE2 or dACE2 proteins. Glycosylated and deglycosylated plant produced ACE2 proteins were purified from N. benthamiana plant using HisPur™ Ni-NTA resin. gACE2: 5 or 10 μg purified gACE2 protein was loaded into well; dACE2: 5 or 10 μg purified dACE2 proteins were loaded into well. Bovine serum albumin (BSA) standards: 1.0, 2.5, and 5.0 μg BSA protein was loaded as a standard protein. (B) Membrane was probed with anti-His6 antibody. gPA83 (plant produced glycosylated protective antigen of B. anthracis, MM ∼100 kDa) and dPA83 (deglycosylated protective antigen of B. anthracis, MM ∼90 kDa) proteins were used as a standard. (C) Membrane was probed with a purified anti-human monoclonal ACE2 antibody (Cat. No. 375801, BioLegend, CA, United States). cACE2: recombinant human ACE2, amino acid (Gln18—Ser740) (Accession: NM_021804.1) was expressed in CHO cells as His tagged protein (cat. no. 792004, BioLegend, CA, United States). The image was taken using a highly sensitive GeneGnome XRQ Chemiluminescence imaging system.