HEK293T cells were electroporated with mRNA encoding B.1351 variant (6P) or B.1.617 variant (6P) proteins using Neon™ Transfection System 10 μL Kit following the standard protocol provided by manufacturer. After 12 h, the cells were collected and resuspended in MACS buffer (D-PBS with 2 mM EDTA and 0.5% BSA). To detect surface-protein expression, the cells were stained with 10 μg/mL ACE2–Fc chimera (Genescript, Z03484) in MACS buffer for 30 min on ice. Thereafter, cells were washed twice in MACS buffer and incubated with PE–anti-human FC antibody (Biolegend, M1310G05) in MACS buffer for 30 min on ice. Live/Dead aqua fixable stain (Invitrogen) were used to assess viability. Data acquisition was performed on BD FACSAria II Cell Sorter (BD). Analysis was performed using FlowJo software.
Pe anti human fc antibody
Manufactured by BioLegend
PE–anti-human FC antibody is a fluorescently labeled antibody that binds to the Fc region of human immunoglobulin molecules. It can be used in flow cytometry and other immunoassays to detect and quantify cells expressing human Fc receptors.
Automatically generated - may contain errors
4 protocols using pe anti human fc antibody
1
SARS-CoV-2 Spike Protein Expression Analysis
2
Omicron Spike Expression in HEK293T Cells
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On day 1, HEK293T cells were seeded at 50% confluence in 24-well plate and mixed with 2 µg Omicron LNP-mRNA. After 16 hours, the cells were collected for flow cytometry. The spike expression on cell surface were detected by staining cells with human ACE2–Fc chimera (Sino Biological, 10108-H02HG) in MACS buffer (D-PBS with 2 mM EDTA and 0.5% BSA) for 20 min on ice. Cells were washed twice after the primary stain and incubated with PE–anti-human Fc antibody (Biolegend, Cat. No. 410708, Clone No. M1310G05, 1:100 dilution) in MACS buffer for 20 min on ice. During secondary antibody staining, live/Dead aqua fixable stain (Invitrogen) was used to assess cell viability. Data was collected on BD FACSAria II Cell Sorter (BD) and analyzed using FlowJo software (version 10.7.2, FlowJo LLC).
3
Spike Protein Expression Profiling
Check if the same lab product or an alternative is used in the 5 most similar protocols
+ Expand
HEK293T cells were electroporated with mRNA encoding SARS, MERS or Delta spikes using Neon™ Transfection System 10 μL Kit following the standard protocol provided by manufacturer. After 12 h, the cells were collected and resuspended. To detect surface-protein expression, the cells were stained with ACE2–Fc chimera (Genscript, Z03484) or DPP4-Fc (Sino Biological, 10688-H01H) in MACS buffer (D-PBS with 2 mM EDTA and 0.5% BSA) for 30 min on ice. Thereafter, cells were washed twice and incubated with PE–anti-human FC antibody (Biolegend, 410708) in MACS buffer for 30 min on ice. Data acquisition was performed on BD FACSAria II Cell Sorter (BD). Analysis was performed using FlowJo software.
4
Omicron Spike Protein Expression Assay
Check if the same lab product or an alternative is used in the 5 most similar protocols
+ Expand
On day 1, HEK293T cells were seeded at 50% confluence in 24-well plate and mixed with 2 μg Omicron LNP-mRNA. After 16 hours, the cells were collected for flow cytometry. The spike expression on cell surface were detected by staining cells with human ACE2–Fc chimera (Sino Biological, 10108-H02HG) in MACS buffer (D-PBS with 2 mM EDTA and 0.5% BSA) for 20 min on ice. Cells were washed twice after the primary stain and incubated with PE–anti-human Fc antibody (Biolegend, 410708) in MACS buffer for 20 min on ice. During secondary antibody staining, live/Dead aqua fixable stain (Invitrogen) was used to assess cell viability. Data was collected on BD FACSAria II Cell Sorter (BD) and analyzed using FlowJo software.
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