5×105-1×106 cells were plated in each well of a round bottom 96-well plate and incubated in the presence of TB10.44-11 peptide (10 μM; New England Peptide). Cells incubated in the presence of αCD3/αCD28 (1 μg/mL; BioLegend) or in the absence of stimuli were used as positive and negative controls, respectively. Cells were incubated for 1 hour at 37° C, at which point GolgiPlug solution (BD Pharmingen, CA, USA) was added to each well for the remaining 4 hours. Cells were collected after the 5 hours stimulation and then surface stained with the antibodies described above, followed by intracellular staining for IFN-γ (clone XMG1.2), TNF (clone MPX6-T22), or granzyme B (clone gb11) using the BD Permwash Kit (BD Pharmingen, CA, USA) as per manufacturer's instructions.
αcd3 αcd28
Manufactured by BioLegend
αCD3/αCD28 is a co-stimulatory reagent used to activate T cells in vitro. It consists of monoclonal antibodies targeting the CD3 and CD28 receptors on the surface of T cells, which are essential for T cell activation and proliferation.
Automatically generated - may contain errors
3 protocols using αcd3 αcd28
1
Antigen-Specific T Cell Activation Assay
2
Quantifying T-cell Cytokine Response
Check if the same lab product or an alternative is used in the 5 most similar protocols
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5 × 105−1 × 106 cells were plated in each well of a round bottom 96-well plate and incubated in the presence of TB10.44–11 peptide (10 μM; New England Peptide). Cells incubated in the presence of αCD3/αCD28 (1 μg/mL; BioLegend) or in the absence of stimuli were used as positive and negative controls, respectively. Cells were incubated for 1 hour at 37 °C, at which point GolgiPlug solution (BD Pharmingen, CA, USA) was added to each well for the remaining 4 hours. Cells were collected after a 5 hour stimulation and then surface stained with the antibodies described above, followed by intracellular staining for IFN-γ (clone XMG1.2), TNF (clone MPX6-T22), using the BD Permwash Kit (BD Pharmingen, CA, USA) as per manufacturer’s instructions.
3
Antigen-Specific T Cell Activation Assay
Check if the same lab product or an alternative is used in the 5 most similar protocols
+ Expand
5×105 cells were plated in each well of a round bottom 96-well plate and incubated in the presence of TB10.44–11 peptide (10 μM; New England Peptide). Incubation in the presence of αCD3/αCD28 (1 μg/mL; BioLegend) or in the absence of stimuli were used as positive and negative controls, respectively. Cells were incubated for 1 h at 37° C, at which point Golgi Stop solution (BD Pharmingen, CA, USA) was added to each well for the remaining 4 h. Cells were collected after the 5 h stimulation and then surface stained with the antibodies described above, followed by intracellular staining for IFNγ (clone) using BD Permwash Kit (BD Pharmingen, CA, USA) as per manufacturer’s instructions.
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