Recombinant SARS-CoV-2 S1 Subunit Protein (RBD), mutant N331Q

C-terminal his-tag

-20°C

Recombinant SARS-CoV-2 S1 subunit protein (RBD) mutant N331Q with C-terminal His-tag, derived from the transfected human HEK293 cells.

12352202

Arg319-Phe541. N-glycosylation site at Asn (N) 331 was mutated to Gln (Q).

His-tag affinity purification by immobilized metal ion affinity chromatography (IMAC)

Figure 1. Deglycosylation of purified recombinant proteins. Purified proteins were untreated (Lane 2) or treated with Protein Deglycosylation Kit under native (Lane 3) or reducing (Lane 4) conditions. Deglycosylation treatment resulted in a mobility shift of the protein to produce one major band at the expected size, thus indicating that the untreated recombinant protein (Lane 2) was glycosylated.Lane 1: Protein standard ladder (kDa)Lane 2: Untreated protein under reducing conditionsLane 3: Treated protein with deglycosylation enzymes under native conditionsLane 4: Treated protein with deglycosylation enzymes under reducing conditions.

Product binding ability was measured by ELISA. The immobilized recombinant human ACE2 protein (Catalog #230-30165, coated at 0.5 ug/ml, 100 ul/well) was incubated with the serial diluted SARS CoV-2 S1 RBD N331 mutant protein (Catalog # 230-30157). The bound mutant protein was detected by mouse anti-S1 RBD monoclonal antibody using ELISA. The calculated EC50 is 138.1 ng/ml.