Rat elisa kit

IL-1β and IL-18 levels in kidney were assessed using a rat ELISA kit (Elabscience Biotechnology Co., Ltd., Wuhan, China) according to the manufacturer's instructions. Caspase-1 activity in HK-2 cells was determined using an enzyme activity assay kit (Beyotime Biotechnology, Shanghai, China), according to the manufacturer's instructions.

The concentration of TGF-β1 was determined from the ovarian tissue using rat ELISA kit obtained from Elabscience Biotechnology Inc. (Wuhan, Hubei, P.R.C., China), in compliance with manufacturer’s guidelines.

Tumor necrotic factor (TNF-α), prostaglandin-E2 (PGE2), interleukin-8 (IL-8), and phosphorylated nuclear factor kappa B (p-NF-κB) detection was conducted according to the manufacturer’s instructions (Elabscience). The stomach tissues (n = 3) were kept in at biofreezer (−80 °C) for storage, homogenize at 15 rpm × 1000 using SilentCrusher M (Heidolph) and the supernatant was collected after centrifugation (at 1350× g for 1 h). The supernatant was then analyzed for TNF-α (Catalog No: E-EL-R0019), PGE2 (Catalog No: E-EL-0034), IL-8 (Catalog No: EKF57830), and p-NF-ƙb (Catalog No: E-EL-R0674) quantification through Elabscience Rat ELISA kit.

Serum insulin concentration was determined following the instructions of the Insulin rat ELISA kit (EMD Millipore, Burlington, MA, USA), absorbance was measured at 450 nm, and the homeostasis model assessment index for insulin resistance (HOMA-IR) was then calculated using the following formula [24 (link)]: $$\mathbf{HOMA}-\mathbf{IR}=\frac{\mathbf{Fasting} \mathbf{insulin}\left(\left(\mathsf{\mu }\mathbf{IU}\right)/\mathbf{mL}\right)\times \mathbf{Fasting} \mathbf{glucose}\left(\mathbf{mg}/\mathbf{dL}\right)}{\mathbf{22.5} \times \mathbf{18}}$$
Serum TG, TC, and HDL–C levels were determined by the enzymatic colorimetric method using reagents obtained from BioMed Diagnostics, Inc. (White City, OR, USA), and absorbance was measured at 546 nm. Serum LDL-C was calculated from TG, TC, and HDL-C concentrations using the following equation [25 ]:
Serum ALT and AST activities were determined using reagents obtained from BioMed Diagnostics, Inc. (USA), and absorbance was measured at 340 nm. Urea and creatinine were determined using reagents obtained from BioMed Diagnostics, Inc. (USA), and absorbance was measured at 570 nm and 510 nm, respectively. Serum leptin was assayed using rat ELISA kit (eBioscience, San Diego, CA, USA), adiponectin and NEFA were assayed using rat ELISA kit (Elabscience, Houston, TX, USA), and serum lipase activity was assayed using colorimetric kit (Spectrum, Alexandria, Egypt). All procedures were performed according to the manufacturer’s instructions.

Levels of IL-17 were assessed in the intestinal tissue homogenate using a commercially available rat ELISA kit (Elabscience Biotechnology Co., Wuhan, China), in accordance with the manufacturer’s instructions.

The BDNF concentration (brain-derived neurotrophic factor) in brain and plasma samples attained from the animals after behavioral studies was determined via an ELISA kit (Elabscience Biotechnology Co., Ltd., Wuhan, Hubei, China). In order to accomplish this, cortices were removed from the brain samples, coagulated with PBS, and centrifuged for 15 minutes at 3000 rpm, and the filtrate was accumulated. According to the manufacturer's instructions, the BDNF concentration in the filtrate of the brain and plasma was finally ascertained through employing its respective rat ELISA kit (Elabscience Biotechnology Co., Ltd., Wuhan, Hubei, China).

Chrysin was purchased from Sigma-Aldrich (St. Louis, MO, USA). A variety of colorimetric kits were obtained from (Bio-Med Diagnostic Inc., White City, OR, USA) for the assay of serum levels of fasting blood glucose (FBG), alanine aminotransferase (ALT), aspartate aminotransferase (AST), bilirubin, triglycerides (TG), total cholesterol (TC), and high-density lipoprotein-cholesterol (HDL-C). ELISA kits for the assay of serum leptin were assayed using a rat ELISA kit (eBioscience, San Diego, CA, USA), adiponectin was assayed using a rat ELISA kit (Elabscience, Houston, TX, USA),and insulin and TNF-α were assayed using rat-specific ELISA kits (Chongqing Biospes Co., Ltd., Chongqing, China, catalog no. BEK1243, and BEK1214, respectively). The hepatic supernatant was used for the determination of phosphorylated-AMPK at Thr172 (P-AMPK), LKB1, and mTOR proteins using a rat-specific ELISA kit (LSBio, Seattle, WA, USA, Cat. No. LS-F36060, LS-F13457, and LS-F17553; respectively), according to the manufacturer’s instructions. Additionally, PGC-1α and mitochondrial transcription factor A (Tfam) proteins were assayed using a rat-specific ELISA kit (MyBioSource, San Diego, CA, USA, Cat No. MBS2706379, and MBS1600609), while nuclear respiratory factor 1 (NRF-1) and SREBP-1c proteins were assayed using rat-specific ELISA kits (Chongqing Biospes Co., Ltd., China, Cat. No. BYEK2318 and BYEK3082).