Biotinylation and Conjugation of SARS-CoV-2 Proteins

Purified and Avi-tagged SARS-CoV-2 Wuhan-Hu-1 RBD or S were biotinylated using the Biotin-Protein Ligase-BIRA kit according to manufacturer’s instructions (Avidity) as described previously^{5 (link)}. Ovalbumin (Sigma-Aldrich, A5503-1G) was biotinylated using the EZ-Link Sulfo-NHS-LC-Biotinylation kit according to the manufacturer’s instructions (Thermo Fisher Scientific). Biotinylated ovalbumin was conjugated to streptavidin-BV711 for human single-cell sorts (BD Biosciences, 563262) or to streptavidin-BB515 for phenotyping (BD Biosciences, 564453). For all human and mouse experiments, RBD was conjugated to streptavidin-PE (BD Biosciences, 554061) and streptavidin-AF647 (BioLegend, 405237)^{5 (link),9}.

Free full text: Click here

Schaefer-Babajew D., Wang Z., Muecksch F., Cho A., Loewe M., Cipolla M., Raspe R., Johnson B., Canis M., DaSilva J., Ramos V., Turroja M., Millard K.G., Schmidt F., Witte L., Dizon J., Shimeliovich I., Yao K.H., Oliveira T.Y., Gazumyan A., Gaebler C., Bieniasz P.D., Hatziioannou T., Caskey M, & Nussenzweig M.C. (2022). Antibody feedback regulates immune memory after SARS-CoV-2 mRNA vaccination. Nature, 613(7945), 735-742.

Publication 2022

Biotin-Protein Ligase-BIRA kit EZ-Link Sulfo-NHS-LC-Biotinylation kit streptavidin-BV711 streptavidin-BB515 streptavidin-PE streptavidin-AF647

Af647 Biotin ligase Biotinylation Cell Human Mouse Ovalbumin Protein Sars cov 2 Streptavidin Sulfo nhs

Corresponding Organization :

Other organizations : Rockefeller University, Howard Hughes Medical Institute

Top 5 similar protocols

Variable analysis

independent variables

Biotinylation of SARS-CoV-2 Wuhan-Hu-1 RBD or S using the Biotin-Protein Ligase-BIRA kit
Biotinylation of ovalbumin using the EZ-Link Sulfo-NHS-LC-Biotinylation kit

dependent variables

Conjugation of biotinylated ovalbumin to streptavidin-BV711 for human single-cell sorts
Conjugation of biotinylated ovalbumin to streptavidin-BB515 for phenotyping
Conjugation of biotinylated RBD to streptavidin-PE and streptavidin-AF647 for human and mouse experiments

control variables

Manufacturer's instructions were followed for biotinylation and conjugation procedures

positive controls

No positive controls specified

negative controls

No negative controls specified

Annotations

Based on most similar protocols

Etiam vel ipsum. Morbi facilisis vestibulum nisl. Praesent cursus laoreet felis. Integer adipiscing pretium orci. Nulla facilisi. Quisque posuere bibendum purus. Nulla quam mauris, cursus eget, convallis ac, molestie non, enim. Aliquam congue. Quisque sagittis nonummy sapien. Proin molestie sem vitae urna. Maecenas lorem.

As authors may omit details in methods from publication, our AI will look for missing critical information across the 5 most similar protocols.

About PubCompare

Our mission is to provide scientists with the largest repository of trustworthy protocols and intelligent analytical tools, thereby offering them extensive information to design robust protocols aimed at minimizing the risk of failures.

We believe that the most crucial aspect is to grant scientists access to a wide range of reliable sources and new useful tools that surpass human capabilities.

However, we trust in allowing scientists to determine how to construct their own protocols based on this information, as they are the experts in their field.

Ready to get started?

Revolutionizing how scientists
search and build protocols!