Quantikine elisa mouse rat porcine canine igf 2 immunoassay kit

The recipient subcutaneous fibrous capsule surrounding the silicon spacer at 6 weeks after pretreatment was procured (GHNF; n = 5, control; n = 6). The recipient subcutaneous fibrous capsules were homogenized using SONICS Vibra-cell (SONICS & MATERIALS, Inc., Newtown, CT, USA) in 5 volumes of RIPA buffer (50 mM Tris–HCl, 150 mM NaCl, 1 mM ethylenediaminetetraacetic acid, 1.0% NP-40, 0.5% sodium deoxycholate, 0.1% sodium dodecyl sulfate) (FUJIFILM Wako Pure Chemical Corporation, Osaka, Japan) supplemented with 1 mM phenylmethylsulfonyl fluoride (PMSF) (SIGMA-ALDRICH, Inc.) and 1.0% Protease Inhibitor Cocktail Set V (FUJIFILM Wako Pure Chemical Corporation). The extract was centrifuged at 10,000 g for 20 min, and then the supernatant was recovered, diluted, aliquoted and frozen at − 20 to − 80 °C. The concentrations of total protein and IGF-2 in the supernatant were measured using a Pierce BCA Protein Assay kit (Thermo Fisher Scientific Inc., Waltham, MA, USA) and a Quantikine ELISA Mouse/Rat/Porcine/Canine IGF-2 Immunoassay Kit (R&D Systems, Minneapolis, MN, USA) according to the manufacturer’s instructions.