SARS-CoV-2 Spike Protein Detection

Cell lysates were prepared as previously described [13 (link),62 (link)]. The following antibodies were used for probing the blots: mouse anti-SARS-CoV/SARS-CoV-2 S (GeneTex), mouse anti-V5 (Thermo Fisher Scientific), rabbit anti-HA (Cell Signaling Technology, Danvers, MA, USA), mouse anti-HIV-1 p24 (NIH/AIDS Reagent Program, ARP-4121), monoclonal anti-β-actin (Sigma-Aldrich), HRP-conjugated anti-rabbit IgG (Cell Signaling Technology) and HRP-conjugated anti-mouse IgG (EMD Millipore). Signals were detected using the enhanced chemiluminescence detection kits Clarity and Clarity Max ECL (Bio-Rad, Hercules, CA, USA) followed by quantitation of bands intensities using the ImageJ software V1.53 (National Institutes of Health; https://imagej.nih.gov/ij/).

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Timilsina U., Ivey E.B., Duffy S., Plianchaisuk A., The Genotype to Phenotype Japan (G2P-Japan) C.o.n.s.o.r.t.i.u.m., Ito J., Sato K, & Stavrou S. (2024). SARS-CoV-2 ORF7a Mutation Found in BF.5 and BF.7 Sublineages Impacts Its Functions. International Journal of Molecular Sciences, 25(4), 2351.

Publication 2024

mouse anti-V5 rabbit anti-HA monoclonal anti-β-actin HRP-conjugated anti-rabbit IgG HRP-conjugated anti-mouse IgG Clarity and Clarity Max ECL

Corresponding Organization : University at Buffalo, State University of New York

Other organizations : The University of Tokyo, National Institute of Infectious Diseases, Kumamoto University, Japan Science and Technology Agency

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Variable analysis

independent variables

Cell lysates preparation method

dependent variables

Protein expression levels of SARS-CoV/SARS-CoV-2 S, V5, HA, HIV-1 p24, and β-actin

control variables

Antibodies used for probing the blots: mouse anti-SARS-CoV/SARS-CoV-2 S, mouse anti-V5, rabbit anti-HA, mouse anti-HIV-1 p24, monoclonal anti-β-actin, HRP-conjugated anti-rabbit IgG, and HRP-conjugated anti-mouse IgG
Signal detection method: enhanced chemiluminescence detection kits Clarity and Clarity Max ECL
Quantitation of band intensities using ImageJ software

positive controls

No positive controls were explicitly mentioned

negative controls

No negative controls were explicitly mentioned

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