Mouse c reactive protein simplestep elisa kit

Serum samples were used for the detection of circulating CRP using Mouse C Reactive Protein SimpleStep ELISA Kit (abcam ab222511, Cambridge, UK), as per the manufacturer instructions. Briefly, 50 µL of standards and samples are added to each well in triplicate, followed by the addition of 50 µL of the antibody mix. After 1 h of incubation at room temperature, the wells are washed multiple times to remove the unbound material. An amount of 100 µL of the TMB development solution is added to each well and during 20 min of incubation is catalyzed by HRP, generating blue coloration. This reaction is then stopped by the addition of 100 µL stop solution, completing any color change from blue to yellow. The signal is generated proportionally to the amount of bound analyte, and the intensity is measured at 450 nm. The serum samples used were diluted 1:400; results are estimated using the standard curve generated using the samples provided by the manufacturer.

The levels of triglyceride (TG) and total cholesterol (T-cholesterol) in the serum were measured using an autoanalyzer (model 7600II; Hitachi, Japan) in a biochemical institutional facility (Korea Non-clinic Test Support Center, Korea). The remaining serum samples were stored in a deep freezer for quantification using ELISA. The frozen serum samples were thawed in ice for 1 h before quantification. The C-reactive protein (CRP) in the serum samples were quantified using ELISA kits (ab222511, Mouse C Reactive Protein Simple Step ELISA Kit, Abcam Inc., USA) according to the protocol provided by the manufacturer.