Tnf α and il 6 elisa kits

Waist circumference and blood pressure were recorded by the standard methods. Waist circumference was measured at the mid-point between the lower border of the last rib and the upper rim of the antero-superior iliac crest. BP was recorded from the left arm in sitting position after 10 min of rest. Two measurements were taken after 5-min interval and an average of two recordings was noted (Ferreira-Hermosillo et al., 2015). Fasting blood sample of 8 ml was taken from the subjects having overnight fast of 8 to 10 h to study different parameters. Blood for DNA extraction was stored at −20°C, and serum was separated and stored at −80°C. Fasting serum glucose and lipid profile including serum TGs, HDL-c and cholesterol were measured by the colorimetric method (Randox Kits, U.K.). Insulin was measured by Human insulin ELISA kit (Elabscience, Germany). Insulin resistance was calculated from fasting serum glucose (mmol/l) and fasting serum insulin (µIU/ml) by HOMA-IR (homeostatic model assessment for insulin resistance) using following formula [20 (link)]:
$\text{HOMA-IR}=\text{Fasting\hspace{0.17em}serum\hspace{0.17em}glucose}\times \text{Fasting\hspace{0.17em}serum\hspace{0.17em}insulin}/22.5$
Serum levels of TNF-α and IL-6 were measured by ELISA using TNF-α and IL-6 ELISA kits (Elabscience, Germany).

The lysates of cerebral tissues were centrifuged at 12,000 rpm for 10 min, and then the contents of TNF-α and IL-6 in the supernatant were measured using the specific ELISA kits based on the manufacturer’s instructions. TNF-α and IL-6 ELISA kits were obtained from Elabscience (Wuhan, China).

A CCK-8 assay kit was purchased from Dojindo Molecular Technologies (Kumamoto, Japan). AST, ALT, BUN, and SCr test kits were supported by the Nanjing Jiancheng Bioengineering Institute (Nanjing, China). TNF-α and IL-6 ELISA kits were purchased from Elabscience Biotechnology (Houston, Texas, United States). A TUNEL assay kit (C10617) was purchased from Thermo Fisher Scientific (Carlsbad, CA, United States). The ROS assay kit and Annexin V-FITC Apoptosis Detection Kit were purchased from Beyotime Biotechnology (Shanghai, China). COX-2 antibody (CST, 12282S) and F4/80 antibody (70076S) were purchased from Cell Signaling Technology (Danvers, MA, United States). BAX (ab32503) and iNOS (ab178945) were obtained from Abcam (Chicago, IL, United States). Bcl-2 antibody (BF9103) and beta-actin (AF7018) were obtained from Affinity Bioscience (Jiangsu, China). The Cyt c antibody (d10933-1-AP) was purchased from Proteintech (Rosemont, IL, United States). Goat anti-Rabbit IgG (H + L) Highly Cross-Adsorbed secondary antibody Alexa Fluor 488 (A11034) and Goat anti-Rabbit IgG (H + L) Highly Cross-Adsorbed secondary antibody Alexa Fluor 555 (A21428) were obtained from Thermo Fisher Scientific (Carlsbad, CA, United States).