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Fluorescent images were captured by a NIKON Eclipse T i. TGF-β1 ELISA was performed using the ELISA kit obtained from R&D Systems.
The concentration of Ang II in plasma was detected using the ELISA Kit (RapidBio Lab, Calabasas, CA, USA).
Soluble BrdU-labelled DNA fragments were quantified using the ELISA kit.
It was then concentrated using 10 kDa concentration columns (Millipore Billerica, MA) and subsequently analyzed using the ELISA kit.
Serum leptin concentrations were measured using enzyme-linked immunoassays (Quantikine R&D Systems, Minneapolis, MN, USA) and adiponectin concentrations using the ELISA kit from B-Bridge International (San Jose, CA, USA).
250 µl was selected as the volume of ACSF as this was the smallest volume compatible with duplicate measurements using the ELISA kit, which requires 100 µl per measurement.
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Epidermal growth factor (EGF) and insulin-like growth factors-1 (IGF-1) in the intestinal content were quantified using the ELISA kits specific for chick (RD Ltd , USA.
IL-18 was measured using the ELISA kits from MBL (Nagoya, Japan).
TNF-α concentrations in some experiments were measured using the ELISA kits (Becton, Dickinson and Company).
Serum levels of zonulin and EGF were assayed using the ELISA kits (Immunodiagnostik, Bensheim, Germany and Boster Biological Technology, Fremont, USA, respectively).
Serum was stored at −80 °C prior to measurement of IL6 using the ELISA-Kit IMMULITE 2000 IL6, Siemens Medical Solutions Diagnostics, Eschborn, Germany.
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