This Corticosterone BioAssay™ ELISA Kit is designed to quantitatively measure Corticosterone present in serum, plasma, urine, extracted dried fecal samples, and tissue culture media samples. This kit measures total corticosterone in serum and plasma and in extracted fecal samples. A corticosterone stock solution is provided to generate a standard curve for the assay and all samples should be read off the standard curve.
Background:Corticosterone (C21H30O4, Kendall’s Compound ‘B’) is a glucocorticoid secreted by the cortex of the adrenal gland. Corticosterone is produced in response to stimulation of the adrenal cortex by ACTH and is the precursor of aldosterone. Corticosterone is a major indicator of stress and is the major stress steroid produced in non-human mammals. Studies involving corticosterone and levels of stress include impairment of long term memory retrieval (1), chronic corticosterone elevation due to dietary restrictions (2) and in response to burn injuries (3). In addition to stress levels, corticosterone is believed to play a decisive role in sleep-wake patterns (4,5).
Sample Type:EDTA Plasma, Heparin Plasma, Serum, Tissue Culture Media and Urine
Sensitivity:24.7pg/ml
Assay Range:78.125-10,000pg/ml
Test Princple:Standards or diluted samples are pipetted into a clear microtiter plate coated with an antibody to capture sheep antibodies. A corticosterone-peroxidase conjugate is added to the standards and samples in the wells. The binding reaction is initiated by the addition of a polyclonal antibody to corticosterone to each well. After an hour incubation the plate is washed and substrate is added. The substrate reacts with the bound corticosterone-peroxidase conjugate. After a short incubation, the reaction is stopped and the intensity of the generated color is detected in a microtiter plate reader capable of measuring 450nm wavelength. The concentration of the corticosterone in the sample is calculated, after making suitable correction for the dilution of the sample, using software available with most plate readers.
Precision:Intra Assay Precision: Four human samples were diluted with Assay Buffer and run in replicates of 24 in an assay. The mean and precision of the calculated Corticosterone concentrations were: Sample 1- 322.5 pg/mL, 2.6% CV Sample 2- 564.3 pg/mL, 2.8% CV Sample 3- 630.3 pg/mL, 3.1% CV Inter Assay Precision: Three human samples were diluted with Assay Buffer and run in replicates of 24 in two assays. The mean and precision of the calculated Corticosterone concentrations were: Sample 1- 732.6 pg/mL, 1.6% CV Sample 2- 402.7 pg/mL, 0.5% CV Sample 3- 594.5 pg/mL, 0.2% CV
