Human mouse tgf beta 1 elisa ready set go

Peripheral blood mononuclear cells (PBMCs) were separated on Histopaque 1077 (Sigma Aldrich), resuspended in complete RPMI 1640, 50 µM β-mercaptoethanol with 10% FCS, and washed twice. Cells were counted, and 1 x 10 6 PBMCs was stimulated with either ConA (Concanavalin A from Canavalia ensiformis, Sigma-Aldrich) 10µg/mL, LPS (Lipopolysaccharides from Salmonella minnesota, Sigma-Aldrich) 10µg/mL, PGN (Peptidoglycan from Staphylococcus aureus, BioChemika, Fluka) 10µg/ml, or with 1 x 10 7 /mL heat killed L. helveticus Lafti® L10. The cells were co-cultured at 37°C and 5% CO 2 for 24 h. After centrifugation at 400 x g for 10 min at 4°C, supernatants were collected and stored frozen at -80 °C. Levels of IFN-γ, IL-10 and IL-4 were measured with commercial ELISA kit (R&D Systems®, USA), according to manufacturers' recommendations, in duplicates.
TGF-β1 level was measured directly from the serum. Samples were diluted 7x and determined by commercial ELISA kit (Human/Mouse TGF beta 1 ELISA Ready-SET-Go, eBioscience, San Diego, USA). Samples were assayed in duplicates; the intra-assay coefficient of variation was 11.06%.