Cort eia kit

CORT EIA was performed using the CORT EIA Kit (Cayman Chemical Company). After buffer, sample and reagent preparation, standards were made according to protocol ranging from 5,000 to 8.2 pg/mL. All reagents, standards, samples and buffers were added to the wells according to a scheme and allowed to incubate for 2 h on a shaker. The plate was then washed 5 times in wash buffer. Right before use, Ellmans reagent was reconstituted and added to the plate and allowed to incubate for 1 h at room temperature in total darkness. The plate was then subsequently read at 405 nm. All values are expressed as pg CORT per mg of feather vane.
The results from the CORT EIA obtained from the Microplate reader (DTX 880, Beckman Coulter) were analyzed using an excel file provided from the company that produced the kit (Cayman Chemical Company). The excel results were then statistically analyzed using SAS.

Blood samples were collected in 1.5 ml ethylenediaminetetraacetic acid-coated microcentrifuge tubes. All samples were kept on ice and later centrifuged at 3000 rpm for 15 min at 4°C. Plasma was transferred to clean 0.2 ml Eppendorf tubes and stored at −80°C until use. Plasma CORT concentrations were measured using a CORT EIA kit (Cayman Chemical, Ann Harbor, MI, USA) as described previously (Regev et al., 2012 (link)).

Plasma CORT was measured as previously described [22 (link)]. Mouse blood was collected at 09:00 into vacutainers containing K3 EDTA. Plasma was isolated via centrifugation at 1000× g for 15 min at 4 °C. The samples were stored at −80 °C until the assay was performed. Quantitation of plasma CORT levels was carried out using the CORT EIA kit (Cayman Chemical, Ann Arbor, MI, USA) according to the manufacturer’s protocol.