5×104 enriched CD4+ T cell from WTKL, C4KL, or WTCL mice were cocultured with 1X105 transduced I-Ab+ M12 cells in triplicate in a 96 well round bottom plate in RPMI with 5% FBS (Omega Scientific), Pen-Strep+L-Glutamine (Cytiva), 10ng/ml ciprofloxacin (Sigma), and 50mM beta-2-mercaptoethanol (Fisher) in the presence of titrating amounts of OVA 323–339 peptide starting at 10µM OVA and a 1:3 titration. The supernatants were collected and assayed for IL-2 concentration by ELISA after 16 hours of co-culture at 37ºC. Anti-mouse IL-2 (clone JES6–1A12, BioLegend) antibody was used to capture IL-2 from the supernatants, and biotin anti-mouse IL-2 (clone JES6–5H4, BioLegend) antibody was used as the secondary antibody. Streptavidin-HRP (BioLegend) and TMB substrate (BioLegend) were also used.
Biotin anti mouse il 2 clone jes6 5h4
Manufactured by BioLegend
Biotin anti-mouse IL-2 (clone JES6–5H4) is a biotin-conjugated monoclonal antibody that binds to mouse interleukin-2 (IL-2). It can be used for the detection and quantification of mouse IL-2 in various immunoassays.
Automatically generated - may contain errors
2 protocols using biotin anti mouse il 2 clone jes6 5h4
1
OVA-Specific CD4+ T Cell Assay
2
Quantitation of IL-2 in CHO I-E^k Co-culture
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For CHO I-Ek co-culture experiments 5 X 104 58α-β- cells were co-cultured with 1 X 105 CHO I-Ek cells and a titration of MCC 88–103 peptide in 96-well flat-bottom plates. Alternatively, 2.5 X 104 58α-β- cells were co-cultured with 1 X 105 M12 cells expressing I-Ek tethered to MCC, T102S or HB in 96-well round-bottom plates. For both assays supernatants were collected after 16 hours of co-culture at 37°C IL-2 was quantitated by ELISA. Anti-mouse IL-2 (clone JES6-1A12, Biolegend) was used as a capture antibody and biotin anti-mouse IL-2 (clone JES6-5H4, Biolegend) was used as the secondary antibody. Streptavidin-HRP and TMB substrate (Biolegend) were used for detection.
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