Rat elisa kit

Rat serum CRP and human serum CRP were measured using ELISA kits (Alpha Diagnostics International, San Antonio, U.S.A.). Blood glucose levels were measured by the glucose oxidase assay (Erba-Lachema, Brno, Czech Republic) using tail vein blood drawn into 5% trichloracetic acid and promptly centrifuged. NEFA levels were determined using an acyl-CoA oxidase-based colorimetric kit (Roche Diagnostics GmbH, Mannheim, Germany). Serum triglyceride concentrations were measured by standard enzymatic methods (Erba-Lachema, Brno, Czech Republic). Serum insulin concentrations were determined using a rat insulin ELISA kit (Mercodia, Uppsala, Sweden). Serum IL6 and TNFα were measured by rat ELISA kits (BioSource International, Inc., Camarillo, U.S.A.).

Blood samples were collected at the time of sacrifice in serum tubes (vacutainer) and were centrifuged at 3000 rpm for 20 min at 4 °C to obtain serum. The serum levels of aspartate aminotransferase (AST), alanine aminotransferase (ALT), and alkaline phosphatase (ALP), total protein (TP), total cholesterol (TC), triglycerides (TG), Conjugated bilirubin, C-reactive protein (CRP), interleukin-6 (IL-6); lactate dehydrogenase (LDH) and Protein induced by vitamin K absence-II (PIVKA-II) were all estimated using the rat ELISA kits obtained by BioSource USA following the instructions and steps contained in the internal kits bulletin.

Serum levels of conjugated bilirubin, total cholesterol (TC), total protein (TP), alanine aminotransferase (ALT), aspartate aminotransferase (AST), triglyceride (TG) and alkaline phosphatase (ALP) were all measured using the rat ELISA kits from BioSource USA and performed in accordance with the instructions and steps outlined in the internal kit’s bulletin.

Biochemical parameters were determined as follows: serum glucose, triacylglycerols and total cholesterol using kits from Erba Lachema (Czech Republic); insulin using an ELISA kit from Mercodia (Sveden); HDL cholesterol and NEFA using kits from Roche Diagnostics (Germany); high-molecular weight (HMW) adiponectin using an ELISA kit (MyBioSource, USA). Plasma concentrations of pro-inflammatory parameters IL-6, MCP-1 and CRP were determined by rat ELISA kits (Bio-Source International, USA; eBioscience, Austria; Alpha Diagnostics International, USA, respectively).

The levels of glucose, triacylglycerols (TG), and total cholesterol (QCA, Barcelona, Spain), and non-esterified free fatty acids (FFAs) (WAKO, Neuss, Germany) were determined in serum samples that were prepared from saphenous vein blood before and after the treatment period (10th and 18th weeks) by enzymatic colorimetric kits. The circulating levels of LDL/VLDL-C and HDL-C and those of insulin were measured from the same samples at the end of the treatment by enzymatic colorimetric kits (Bioassay systems, California, CA, USA) and while using a rat/mouse ELISA kit (Millipore, Barcelona, Spain), respectively. Insulin resistance and sensitivity were assessed while using the HOMA-IR and R-QUICKI indices with the following formulas: (Glucose x Insulin)/22.5 and 1/[log insulin (µU/mL) + log glucose (mg/dL) + log FFA (mmol/l)], respectively.
From serum samples that were obtained from cardiac puncture at the end of the study, we measured vascular cell and intercellular adhesion molecule 1 (VCAM-1 and ICAM-1) and serum monocyte chemoattractant protein 1 (MCP-1) while using rat ELISA kits (Thermo Scientific, Illinois, USA). The Neuraminidase (NA) activity was measured with the Amplex^TM Red Neuraminidase Assay Kit (Thermo Scientific, Illinois, IL, USA).