CD4+ T cells were purified from inguinal lymph nodes (draining immunization site) of Batf3−/− mice after induction of a DTH reaction using the CD4+ T cell MACS separation kit (Miltenyi Biotech). For DC isolation, spleens from untreated C57BL/6 wild-type mice were isolated and digested with Liberase TL (Roche, Indianapolis, IN, USA) in the presence of DNAse I (Roche) at 37 °C. CD11c+ cells were purified from the digested cell suspensions using a CD11c+ DC MACS separation kit (Miltenyi Biotech). For in vitro restimulation, 1×104 CD11c+ cells were pulsed with 0.5 mg/ml OVA protein and cocultured with 1×105 CD4+ T cells for 48 hours. Alternatively, 5×105 cells from the inguinal lymph nodes of MLN-resected mice were restimulated with OVA protein. Subsequently, protein levels of IFNγ were measured in culture supernatant by mouse ELISA set kit (Biolegend, San Diego, CA, USA) according to the manufacturer's instructions. The sensitivity of the assay was 4 pg/ml.
Mouse elisa set kit
Manufactured by BioLegend
Sourced in United States
The Mouse ELISA set kit is a laboratory equipment designed for the quantitative measurement of target analytes in mouse samples. This kit provides the necessary components to perform enzyme-linked immunosorbent assay (ELISA) experiments, including pre-coated plates, detection antibodies, and other essential reagents.
Automatically generated - may contain errors
2 protocols using mouse elisa set kit
1
Purification and Stimulation of CD4+ T Cells
2
Purification and Stimulation of CD4+ T Cells
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CD4+ T cells were purified from inguinal lymph nodes (draining immunization site) of Batf3−/− mice after induction of a DTH reaction using the CD4+ T cell MACS separation kit (Miltenyi Biotech). For DC isolation, spleens from untreated C57BL/6 wild-type mice were isolated and digested with Liberase TL (Roche, Indianapolis, IN, USA) in the presence of DNAse I (Roche) at 37 °C. CD11c+ cells were purified from the digested cell suspensions using a CD11c+ DC MACS separation kit (Miltenyi Biotech). For in vitro restimulation, 1×104 CD11c+ cells were pulsed with 0.5 mg/ml OVA protein and cocultured with 1×105 CD4+ T cells for 48 hours. Alternatively, 5×105 cells from the inguinal lymph nodes of MLN-resected mice were restimulated with OVA protein. Subsequently, protein levels of IFNγ were measured in culture supernatant by mouse ELISA set kit (Biolegend, San Diego, CA, USA) according to the manufacturer's instructions. The sensitivity of the assay was 4 pg/ml.
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