Anti-SARS-CoV-2 Spike Glycoprotein RBD Antibody [5G8] (A270557)

Immunofluorescent analysis of HEK293 cells transfected with pCI-Neo-Mod vector including SARS-CoV-2 Spike Glycoprotein RBD cDNA, and stained with Anti-SARS-CoV-2 Spike Glycoprotein RBD Antibody [5G8] (A270557), at a dilution of 1:1,000, in red. Nuclear DNA is visualised in blue using Hoechst staining. Anti-SARS-CoV-2 Spike Glycoprotein RBD Antibody [5G8] (A270557) reveals expression of SARS-CoV-2 Spike Glycoprotein RBD protein only in transfected cells. DAPI reveals the nuclear DNA of both transfected and non-transfected cells.

Western blot analysis of HEK293 cell lysates using Anti-SARS-CoV-2 Spike Glycoprotein RBD Antibody [5G8] (A270557), at a dilution of 1:3,000, in green. The lanes contain: [Lane 1] protein standard, [Lane 2] non-transfected cells, [Lane 3] cells transfected with pCI-Neo-Mod containing the SARS-CoV-2 Spike glycoprotein RBD cDNA, and [Lane 4] cells transfected with a pCI-Neo-GFP vector expression construct containing the SARS-CoV-2 Spike Glycoprotein RBD cDNA. The band at 25 kDa in the transfected cells demonstrates expression of SARS-CoV-2 Spike Glycoprotein, and the band at about 50 kDa corresponds to the GFP-SARS-CoV-2 Spike Glycoprotein fusion protein. The same blot was simultaneously probed with Anti-HSP60 Antibody (A85437), at a dilution of 1:5,000, in red, revealing a single band at 60 kDa in both transfected and non-transfected cells, acting as a positive loading control.

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 [5G8] (A270557), Anti-SARS-CoV-2 Spike Glycoprotein RBD Antibody [2G1] (A270550) and Anti-SARS-CoV-2 Spike Glycoprotein RBD Antibody (A270592) 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.

(Left) Western blot analysis of HEK293 cell lysates using Anti-SARS-CoV-2 Spike Glycoprotein RBD Antibody [5G8] (A270557) at a dilution of 1:3,000 (green). The lanes contain: [Lane 1] protein standard, [Lane 2] non-transfected cells, [Lane 3] cells transfected with pCI-Neo-Mod containing the SARS-CoV2-bd cDNA, and [Lane 4] cells transfected with pCI-Neo-GFP vector expression construct containing containing the SARS-CoV2-bd cDNA. The band at 25kDa mark in the transfected cells demonstrates expression of SARS-CoV-bd protein, and the band at about 50kDa corresponds to a GFP-SARS-CoV2-bd fusion protein. The same blot was simultaneously probed with Anti-HSP60 Antibody (A85437) at a dilution of 1:5,000 (red) revealing a single band at 60kDa in both transfected and non-transfected cells. (Right) Western Blot of full length recombinant SARS-CoV2 S-protein expressed in HEK293 cells using product 10561-CV from R&D Systems. Lane 6 shows a loading of 1µg and lane 7 is 100ng. On longer exposure of the blot the antibody could readily detect 10ng of the S-protein. Lanes 1 and 5 are molecular weight standards of indicated size.