ELISA and Real-Time PCR
Collected blood samples were allowed to clot and centrifuged at 10,000 rpm for 10 min at 4°C to separate out serum. Standard sandwich ELISA was performed to assess IgE levels (BD biosciences). For real-time PCR, RNA was isolated from snap-frozen proximal colon lysates using TRIzol extraction according to the manufacturers’ instruction. Real-time PCR was performed on cDNA generated by using a Superscript II reverse transcriptase kit (Life Technologies) with SYBR Green master mix (Applied Biosystems) and commercially available primer sets (QIAGEN QuantiTect). Samples were run on the ViiA7 real time PCR systems (Applied Biosystems -Life Technologies).