Wt and Pla2g5-null mice sorted ILC2s were obtained as described above and rested for 40h with 10ng/mL rIL-2 and rIL-7 in a 96 well round bottom plates (40000 cells per well). After changing to fresh media, ILC2s were stimulated with 30ng/mL rIL-33 or 30ng/mL rIL-33, 200 μM Linoleic Acid, 200 μM Oleic Acid for 8h. Supernatants were then collected. IL-5 ELISA (R&D Systems, M5000) was performed as per manufacturer protocol. For these experiments, during lung homogenization, Dispase (Gibco, Life Technologies, NY) was added (2U/ml) to increase the yield for ILC2s.
Il 5 elisa
Manufactured by R&D Systems
The IL-5 ELISA is a quantitative sandwich enzyme-linked immunosorbent assay designed for the measurement of human interleukin-5 levels in cell culture supernatants, serum, and plasma samples.
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Lab products found in correlation
3 protocols using il 5 elisa
1
Measuring IL-5 Production in ILC2s
2
Quantification of IL-5 in Lung Cell Cultures
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Single cell suspensions from the lungs were prepared as described above, and 5 x 105 cells were seeded in 0.1 ml cell cultures with 25 μg of OVA (Sigma, GradeV) in IMDM medium (10% FCS, 1% penicillin/streptomycin), followed by 72 hrs of incubation at 37°C. IL-5 in the supernatants of the OVA-restimulated cells was quantitated by an IL-5 ELISA (R&D Systems, Minneapolis, MN).
3
Measuring IL-5 Production in ILC2s
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+ Expand
Wt and Pla2g5-null mice sorted ILC2s were obtained as described above and rested for 40h with 10ng/mL rIL-2 and rIL-7 in a 96 well round bottom plates (40000 cells per well). After changing to fresh media, ILC2s were stimulated with 30ng/mL rIL-33 or 30ng/mL rIL-33, 200 μM Linoleic Acid, 200 μM Oleic Acid for 8h. Supernatants were then collected. IL-5 ELISA (R&D Systems, M5000) was performed as per manufacturer protocol. For these experiments, during lung homogenization, Dispase (Gibco, Life Technologies, NY) was added (2U/ml) to increase the yield for ILC2s.
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