Recombinant SARS-CoV-2 S1 Subunit Protein RBD, R346K/E484K/N501Y mutant

Proteins|Recombinant Proteins|Viral Proteins|COVID-19|Proteins|Spike Protein|Spike Protein Variants

Blue ice

Recombinant protein product has a calculated molecular mass of ˆ¼25 kDa. Due to the abundant glycosylation, it migrates as approximately ˆ¼30 kDa protein bands in SDS-PAGE under DTT, beta-mercaptoethanol reducing conditions. After deglycosylation under native and denature conditions, the protein presented as one reduced ˆ¼25 kDa band.

Supplied as a 0.2 um filtered solution in PBS (pH 7.4)

Spike protein, S Protein, S1 Subunit, Host Cell Receptor Binding Domain (RBD)

Lot specific (see the label on the vial), determined by BCA protein assay (lot specific)

Figure 1. Deglycosylation analysis of purified recombinant proteins. The same amount of purified proteins were untreated (Lane 2) or treated with protein deglycosylation enzymes under native (Lane 3) or reducing (Lane 4) conditions. Deglycosylation treatment resulted in a mobility shift of the protein to produce one reduced 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 Lane 3: Treated protein with deglycosylation enzymes under native conditions. Lane 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 R346K/E484K/N501Y mutant protein (Catalog #230-30213). The bound mutant protein was detected by mouse anti-S1 RBD monoclonal antibody using ELISA. The calculated EC50 is 39.48 ng/ml.