Anti-SARS-CoV-2 Spike Glycoprotein RBD Antibody [2G1] (A270550)

Left Panel: HEK293 cells were transfected with DNA encoding the SARS-CoV-2 Spike Glycoprotein RBD for ACE2 which had been inserted into pCI-Neo-Mod or pCI-Neo-GFP eukaryotic expression vectors, such that the cells express either the insert only or the insert fused with GFP. Lane 2 shows a crude homogenate of untransfected control cells, lane 3 shows a homogenate of cells expressing SARS-CoV-2 Spike Glycoprotein RBD, and lane 4 shows a homogenate of cells expressing GFP-SARS-CoV-2 Spike Glycoprotein RBD fusion. The Anti-SARS-CoV-2 Spike Glycoprotein RBD Antibody [2G1] (A270550) was used at a dilution of 1:3,000 and produces clean and specific staining of bands of the expected size as shown in green. The blot was also stained with Anti-Heat Shock Protein 60 Antibody (A85437), at a dilution of 1:5,000 in red.

Right Panel: Blot of full-length recombinant SARS-CoV-2 Spike Glycoprotein expressed in HEK293 cells, product 10561-CV, obtained from R&D Systems. Lane 6 shows a loading of 1 µg and lane 7 is 100 ng. 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.

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Immunoflourescent analysis of HEK293 cells transfected with DNA encoding the S-protein segment in SARS-CoV-2 Spike Glycoprotein RBD which was inserted in the pCI-Neo-Mod expression vector. The Anti-SARS-CoV-2 Spike Glycoprotein RBD Antibody [2G1] (A270550), in green, produces clean and specific staining of transfected cells which stain identically with Anti-SARS-CoV-2 Spike Glycoprotein RBD Antibody (A270592), a rabbit polyclonal antibody to the same immunogen. The nuclei of transfected and untransfected cells are shown in blue with DAPI DNA stain.

Immunofluorescent analysis of HEK293 cells transfected with a SARS-CoV-2 Spike Glycoprotein RBD for ACE2 construct under high magnification, stained with Anti-SARS-CoV-2 Spike Glycoprotein RBD Antibody [2G1] (A270550), at a dilution of 1:1,000, in green. Cells were co-stained with Anti-Heat Shock Protein 60 Antibody (A85437), at a dilution of 1:2,000, in red. The nuclear DNA is visualised in blue using Hoechst staining. Anti-SARS-CoV-2 Spike Glycoprotein RBD Antibody [2G1] (A270550) reveals overexpression of SARS-CoV-2 Spike Glycoprotein RBD presumably in lysosomes only in transfected cells, while the Anti-Heat Shock Protein 60 Antibody (A85437) labels mitochondria in all cells.