As described previously [28 (link)], splenocytes were incubated with SARS-CoV-2 S peptide pools (1 μg/mL, Miltenyi Biotec, Auburn, CA, USA) for 5 h in the presence of BD GolgiPlug (BD Biosciences, San Jose, CA, USA). Briefly, cells were stained with antibodies for CD4 or CD8, fixed in 2% paraformaldehyde, and permeabilized with 0.5% saponin before adding anti-IFN-γ, anti-TNF-α, or control rat IgG1 (e-Biosciences). Samples were processed with a C6 Flow Cytometer instrument. Dead cells were excluded based on forward and side light scatter. Data were analyzed with a CFlow Plus Flow Cytometer (BD Biosciences, San Jose, CA, USA).
Control rat igg1
Manufactured by Thermo Fisher Scientific
Control rat IgG1 is a laboratory reagent used as a control in various immunological assays. It is a purified immunoglobulin G (IgG) antibody derived from rat serum. The primary function of this product is to serve as a reference or control sample to establish baseline or background levels in experiments involving rat-derived antibodies or proteins.
Automatically generated - may contain errors
3 protocols using control rat igg1
1
Quantifying SARS-CoV-2 Specific T Cell Responses
2
Zika Virus-Induced IFN-γ in T Cells
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Splenocytes (2.5 × 106) were incubated with 0.1 × 106 FFU live ZIKV-FSS13025 for 24 h. BD GolgiPlug (BD Bioscience) was added to block protein transport at the final 6 h of incubation. Cells were stained with antibodies for CD3, CD4, or CD8 fixed in 2% paraformaldehyde and permeabilized with 0.5% saponin before adding anti-IFN-γ, or control rat IgG1 (e-Biosciences). Samples were processed with a C6 Flow Cytometer instrument. Dead cells were excluded on the basis of forwarding and side light scatter. Data were analyzed with a CFlow Plus Flow Cytometer (BD Biosciences).
3
SARS-CoV-2 T cell response assay
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Splenocytes or lung leukocytes were incubated with SARS-CoV-2 S peptide pools (1μg/ml, Miltenyi Biotec) for 24 h. BD GolgiPlug (BD Bioscience) was added to block protein transport at the final 6 h of incubation. Cells were stained with antibodies for CD3, CD4, or CD8, fixed in 2% paraformaldehyde, and permeabilized with 0.5% saponin before adding anti-IFN-γ, or control rat IgG1 (e-Biosciences). Samples were processed with a C6 Flow Cytometer instrument. Dead cells were excluded based on forward and side light scatter. Data were analyzed with a CFlow Plus Flow Cytometer (BD Biosciences).
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