B cell-specific tetramers were prepared as described55 (link). A biotinylated form of recombinant S1 and RBD proteins (BioLegend) was tetramerized by the addition of PE-conjugated or BV421-conjugated streptavidin (BioLegend) and used for B cell tetramer staining assays. Briefly, SA-PE or SA-BV21 was added in an amount equal to 1/5 of the monomer substrate amount. SA was added in 5 equal portions to the monomer and incubated each time at 4 °C for 20 min on a shaker. Protease inhibitor was added to the tetramers at a 1x final concentration (Sigma). The tetramers were filled up to 100 µl with 0.1% BSA in PBS and stored at 4 °C.
Jacobsen E.M., Fabricius D., Class M., Topfstedt F., Lorenzetti R., Janowska I., Schmidt F., Staniek J., Zernickel M., Stamminger T., Dietz A.N., Zellmer A., Hecht M., Rauch P., Blum C., Ludwig C., Jahrsdörfer B., Schrezenmeier H., Heeg M., Mayer B., Seidel A., Groß R., Münch J., Kirchhoff F., Bode S.F., Strauss G., Renk H., Elling R., Stich M., Voll R.E., Tönshof B., Franz A.R., Henneke P., Debatin K.M., Rizzi M, & Janda A. (2022). High antibody levels and reduced cellular response in children up to one year after SARS-CoV-2 infection. Nature Communications, 13, 7315.
Corresponding Organization : University of Freiburg
Other organizations :
Universität Ulm, University Medical Center Freiburg, University Children's Hospital Tübingen, University Hospital Heidelberg, Heidelberg University
Biotinylated form of recombinant S1 and RBD proteins
PE-conjugated or BV421-conjugated streptavidin
dependent variables
B cell tetramer staining assays
control variables
Protease inhibitor added to the tetramers at a 1x final concentration
Tetramers stored at 4 °C
controls
Positive control: Not explicitly mentioned.
Negative control: Not explicitly mentioned.
Annotations
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