RBD protein expressed with AviTag was purchased from GenScript. Site-specific biotinylation of the AviTag was performed using BirA Biotin-Protein Ligase Reaction kit (Avidity). Next, unconjugated biotin was removed using Zeba spin desalting columns, 7K MWCO (ThermoFisher). The quantification of reacted biotin was performed using the Pierce Biotin Quantification Kit (ThermoFisher). Biotinylated RBD was incubated with either streptavidin-conjugated PE (Biolegend) or streptavidin-conjugated APC fluorophores (Biolegend) for 20 min on ice at a molar ratio of 4:1 of biotin to streptavidin. FITC-labelled streptavidin (Biolegend) was reacted with excess free biotin to form a non-RBD-specific streptavidin probe as a control. Tetramer formation was confirmed using SDS-PAGE gel. Cells were stained for flow cytometry with all three streptavidin probes at the same time as other fluorescent surface markers at a volumetric ratio of 1:100 for RBD-streptavidin-PE and 1:200 for RBD-streptavidin-APC and biotin-streptavidin-FITC.
Streptavidin conjugated apc fluorophores
Manufactured by BioLegend
Streptavidin-conjugated APC fluorophores are a type of detection reagent used in flow cytometry and other bioassays. Streptavidin is a protein that binds to the small molecule biotin with high affinity. APC is a fluorescent dye that can be detected using flow cytometry instrumentation. The streptavidin-APC conjugate allows for the detection of biotinylated molecules in a sample.
Automatically generated - may contain errors
Lab products found in correlation
Pierce biotin quantification kit, by Thermo Fisher Scientific (2 mentions)
Pe conjugated streptavidin, by BioLegend (2 mentions)
Avitag, by GenScript (2 mentions)
Avitag, by Avidity (2 mentions)
Bira biotin protein ligase reaction kit, by Avidity (2 mentions)
Fitc conjugated streptavidin, by BioLegend (1 mentions)
2 protocols using streptavidin conjugated apc fluorophores
1
Biotinylated RBD Labeling for Flow Cytometry
2
Biotinylated RBD Tetramer Assay
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RBD protein expressed with AviTag was purchased from GenScript. Site-specific biotinylation of the AviTag was performed using BirA Biotin-Protein Ligase Reaction kit (Avidity). Next, unconjugated biotin was removed using Zeba spin desalting columns, 7K MWCO (ThermoFisher). The quantification of reacted biotin was performed using the Pierce Biotin Quantification Kit (ThermoFisher). Biotinylated RBD was incubated with either streptavidin-conjugated PE (Biolegend) or streptavidin-conjugated APC fluorophores (Biolegend) for 20 min on ice at a molar ratio of 4:1 of biotin to streptavidin. Streptavidin-conjugated FITC (BioLegend) was reacted with excess free biotin to form a non-RBD-specific streptavidin probe as a control. Tetramer formation was confirmed using SDS-PAGE gel. Cells were stained for flow cytometry with all three streptavidin probes at the same time as other fluorescent surface markers at a volumetric ratio of 1:100 for RBD-streptavidin-PE and 1:200 for RBD-streptavidin-APC and biotin-streptavidin-FITC.
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