Ige elisa kit

TM E/S antigen-specific serum IgG1 and IgG2c ELISAs were carried out as described [15 (link)]. Briefly, serum was added to ELISA plates that had been coated overnight with TM E/S antigen (5ug/ml), washed and blocked in 5% fetal calf serum. Following a 2h 37°C incubation, plates were sequentially washed, incubated for 1 hour at room temperature with secondary antibodies (BD Biosciences: biotinylated anti-mouse IgG1, clone A85-1 or biotinylated anti-mouse IgG2c, clone R19-15), washed and incubated with streptavidin HRP (BD Pharmingen) for 30 minutes at room temperature and finally detected with TMB substrate (eBioscience). Enzymatic reactions were stopped with 1N H₂SO₄ and OD450 measured on a Spectramax (Molecular Devices) spectrophotometer.
IgE ELISA was carried out by using the IgE ELISA kit from Thermofisher according to manufacturer’s instructions. OD values were measured using a Spectrophotometer (Versamax).
IFNγ ELISA was carried out with the Mouse IFN gamma ELISA Ready-SET-Go! Kit (eBioscience, cat #88–7314) according to manufacturer’s instructions. TSLP (Mouse TSLP Quantikine ELISA kit, R&D systems) and IL13, IL10 and IL4 were measured using a Milliplex Map kit (Millipore) according to manufacturer’s instructions.