Anti-SARS-CoV-2 Spike Glycoprotein RBD Antibody (A270592)

Immunofluorescent analysis of HEK293 cells transfected with pCI-Neo-Mod vector encoding SARS-CoV-2 Spike Glycoprotein RBD, corresponding to amino acids 308-541 of the S-protein sequence in the Wuhan-Hu-1 complete genome, stained with Anti-SARS-CoV-2 Spike Glycoprotein RBD Antibody (A270592), at a dilution of 1:3,000, in red, and co-stained with Anti-beta Tubulin Antibody [4E4] (A85429), at a dilution of 1:5,000, in green. The nuclear DNA is visualised in blue using Hoechst staining. Anti-SARS-CoV-2 Spike Glycoprotein RBD Antibody (A270592) cleanly reveals expression of the SARS-CoV-2 Spike Glycoprotein RBD only in transfected cells. Anti-beta Tubulin Antibody [4E4] (A85429) produces strong staining of microtubules in the cytoplasm in both transfected and non-transfected cells.

Left: Western blot analysis of HEK293 cell lysates using Anti-SARS-CoV-2 Spike Glycoprotein RBD Antibody (A270592), at a dilution of 1:2,000, in green. The lanes contain: [Lane 1] protein standards (red), [Lane 2] non-transfected cells, [Lane 3] cells transfected with pCI-Neo-Mod vector encoding the SARS-CoV-2 Spike Glycoprotein RBD, and [Lane 4] cells transfected with pCI-Neo-GFP vector encoding the SARS-CoV-2 Spike Glycoprotein RBD. The band at 25 kDa in Lane 3 demonstrates expression of SARS-CoV-2 Spike Glycoprotein RBD, and the band at approximately 50 kDa in Lane 4 corresponds to the GFP-SARS-CoV-2 Spike Glycoprotein RBD fusion protein. The same blot was simultaneously probed with Anti-beta Tubulin Antibody [4E4] (A85429), at a dilution of 1:5,000, in red, which reveals a single band at 50 kDa in both transfected and non-transfected cells, so acts as a positive loading control. Right: Western blot analysis of full length recombinant SARS-CoV-2 Spike Glycoprotein (10561-CV from R&D Systems) using Anti-SARS-CoV-2 Spike Glycoprotein RBD Antibody (A270592), in green. The lanes contain: [Lane 5] protein standards (red), [Lane 6] 1 µg of protein, and [Lane 7] 100 ng of protein. On longer exposure of the blot, Anti-SARS-CoV-2 Spike Glycoprotein RBD Antibody (A270592) could readily detect 10 ng of the Spike glycoprotein.

Vero cells are a cell line derived derived from monkey kidney and express an ACE2 receptor similar to that found in humans, and are therefore susceptible to infection by patient derived SARS-CoV2 virus. The cells were grown in culture, fixed, and stained with the DNA dye DAPI, revealing cell nuclei (control, top left), The remaining three panels show staining of similar cells following infection with SARS-CoV2 virus. Cells were then reacted with the cell binding domain of the SARS-CoV2 spike protein, namely Anti-SARS-CoV-2 Spike Glycoprotein RBD Antibody (A270592), Anti-SARS-CoV-2 Spike Glycoprotein RBD Antibody [2G1] (AA270550) and Anti-SARS-CoV-2 Spike Glycoprotein RBD Antibody [5G8] (A270557) as indicated. Antibody binding was revealed with appropriate ALEXA green (top right, bottom left) or red (bottom right) secondary antibodies. The virus accumulates in the cytoplasm of infected cells.