Fresh peripheral blood mononuclear cells (PBMCs) were used in all ELISpot assays using the ELISpot IFN-γ kit (AID). Briefly, fresh PBMCs (2 × 105 cells in 50 μL) were placed in duplicate wells and stimulated with 50 μL SARS-CoV-2 peptide pools (S-complete, Miltenyi Biotech) (2 μg/mL per peptide). Phytohemagglutinin (PHA) was used as a positive control, and 4Cell Nutri-T-medium was used as negative control. After 16–20 h at 37 °C, 5% CO2, and 95% humidity, cells were removed and secreted IFN-γ was detected by adding an alkaline phosphatase-conjugated secondary antibody for 2 h. The plates were developed using BCIP/NBT substrate according to the manufacturer’s instructions. The ELISpot plates were scanned using an AID ELISpot reader. Nonspecific background (mean SFU from the negative control wells) was subtracted from the experimental readings.
Sars cov 2 peptide pools s complete
Manufactured by Miltenyi Biotec
The SARS-CoV-2 peptide pools (S-complete) are a set of overlapping peptides covering the full-length SARS-CoV-2 spike (S) protein sequence. These peptide pools are designed for use in T cell immune response studies related to SARS-CoV-2 infection.
Automatically generated - may contain errors
Lab products found in correlation
2 protocols using sars cov 2 peptide pools s complete
1
SARS-CoV-2 Specific T-Cell Response
2
SARS-CoV-2 T-cell Evaluation by ELISpot
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Fresh PBMCs were used in all ELISpot assays using the Elispot IFNγ kit (AID). Briefly, fresh PBMCs were added in duplicate wells at 2x105 (link) cells in 50 μl per well and stimulated with 50 μl of SARS-CoV-2 peptide pools (S-complete, Miltenyi Biotech) (2 μg/ml per peptide). 4Cell Nutri-T-Medium was used as a negative control and phytohaemagglutinin was used as a positive control. After 16–20 h at 37 °C, 5% CO2, 95% humidity, cells were removed and secreted IFNγ was detected by adding alkaline phosphatase-conjugated secondary antibody for 2 h. The plates were developed using BCIP/NBT substrate according to the manufacturer’s instructions. ELISpot plates were scanned on an AID ELISpot Reader. The unspecific background (mean spot-forming units from negative control wells) was subtracted from experimental readings. It was only possible to perform the T-cell evaluation test over 2 days, so only the patients who agreed to participate during these days received a T-cell evaluation.
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