Miltenyi Biotec-tested panel 42 (MBTP 42)
This application protocol describes the analysis of SARS-CoV-2–specific B cells using recombinant SARS-CoV-2 spike proteins and flow cytometry.
For the preparation of spike-tetramer solutions, refer to the application protocol Analysis of SARS-CoV-2–specific B cells using recombinant SARS-CoV-2 proteins and to the data sheet of the SARS-CoV-2 Spike B Cell Analysis Kit, human.
For preparation of the staining cocktail from the SARS-CoV-2 Spike B Cell Analysis Kit, human, 2 μL of each fluorochrome-conjugated antibody, 5 μL of 7-AAD Staining Solution, 5 μL of spike‑tetramer-PE, and 10 μL of spike-tetramer-PE-Vio 770 were mixed and filled up to 100 μL with PEB buffer for each test (5×106 B cells or 107 PBMCs).
▲Note: CD19+ cells can be enriched from a single-cell suspension of PBMCs using the REAlease CD19 MicroBead Kit, human according to the data sheet.
|Specificity||Clone||Purpose||Fluorochrome||Detection filter (nm) (laser)|
|CD19||LT19||Pan B cells||APC-Vio® 770||750 LP (red)|
|CD27||M-T271||Memory B cells||Vio Bright FITC||525/50 (violet)|
|IgA||IS11-8E10||Subset of memory B cells||VioGreen™||525/50 (violet)|
|IgG||IS11-3B2.2.3||Subset of memory B cells||VioBlue®||450/50 (violet)|
|IgM||PJ2-22H3||Subset of memory B cells||APC||655–730 (red)|
|Streptavidin||-||Spike-tetramer||PE-Vio 770||750 LP (blue)|
|7-AAD Staining Solution||-||Viability||-||655-730 (blue)|
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