Rat tnf α elisa kit

The following reagents and kits were used in the present study: Rabbit polyclonal anti-tyrosine hydroxylase (TH; cat. no. 2792; Cell Signaling Technology, Inc., Boston, MA, USA), rabbit polyclonal anti-OX-42 (cat. no. orb11009; Biorbyt, Cambridge, UK), hydroethidine (Molecular Probes, Eugene, OR, USA), LPS (Sigma-Aldrich, St. Louis, MO, USA), heparin (Qianhong Bio-pharma Co., Ltd., Changzhou, China), SST (ProSpec, East Brunswick, NJ, USA), biotinylated goat anti-rabbit secondary antibody (cat. no. A0277) and horseradish peroxidase (HRP)-labeled streptavidin (Beyotime, Shanghai, China), bicinchoninic acid (BCA) kit (Beyotime), Rat TNF-α ELISA kit, Rat IL-1β ELISA kit and Prostaglandin E2 ELISA kit (PGE2; USCN, Wuhan, Hubei, China).

Testicular levels of TNF-α was estimated as described in previous studies using rat TNF-α ELISA kit (USCN Life Science, Houston, TX, USA) [17 (link), 26 (link)] based on a sandwich enzyme immunoassay. The manufacturer’s instructions were followed. Standards or samples were added to the microtiter plate wells with a biotin-conjugated antibody specific to TNF-α. Afterwards, avidin conjugated to HRP was added and incubated. After the addition of TMB substrate solution, wells that contained TNF-α, biotin-conjugated antibody and enzyme-conjugated avidin would display a change in color. The enzyme-substrate reaction was ended by the addition of sulfuric acid and the color change was determined at 450 nm.