Mouse tgfβ1 elisa kit

Extracts were prepared from snap frozen liver by homogenisation in lysis buffer (Tris-HCl 50 mmol/L, NaCl 150 mmol/L, EDTA 1 mmol/L, 1% Triton X-100, 0.5% Tween-20, 0.1% SDS) containing a protease inhibitor cocktail (#11836170, Roche Diagnostics, Mannheim, Germany) followed by centrifugation at 14000 × g for 15 min at 4 °C. Supernatants were collected and activated with acetic acid/urea prior to analysis. TGFβ1 content of liver protein extracts were measured using a mouse TGFβ1 ELISA kit (R and D Systems Inc, Minneapolis, MN, United States). Plates were read using the Bio-Rad microplate reader at 450 nm and TGFβ1 concentrations were calculated from the standard curve by the plate reader software.

The amounts of TGF‐β1 in the culture supernatants were determined using the mouse TGF‐β1 ELISA kit (R&D Systems, Minneapolis, MI, USA) according to the manufacturer's instructions. After LLC cells (5 × 10⁵ cells/6 cm dish) were cultured with 3 mL of PANSERIN 401 serum free medium (PAN Biotech, Passau, Germany) for 24 h, media were collected. For determination of the total TGF‐β1 concentrations, media were treated according to the method of an acidification procedure prior to the ELISA assay.