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.Īllow the kit reagents to come to room temperature for 30 minutes. The substrate reacts with the bound corticosterone-peroxidase conjugate.Īfter 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 binding reaction is initiated by the addition of a polyclonal antibody to corticosterone to each well.Īfter an hour incubation the plate is washed and substrate is added. Standards or diluted samples are pipetted into a clear microtiter plate coated with an antibody to capture sheep antibodies.Ī corticosterone-peroxidase conjugate is added to the standards and samples in the wells. Please choose the standard range that fits your sample concentrations most appropriately. We provide protocols on page 8 to prepare assay standards from 5,000 to 78.125 pg/mL or from 10,000 to 78.125 pg/mL. This kit measures total corticosterone in serum and plasma and in extracted fecal samples.Ī corticosterone stock solution is provided to generate a standard curve for the assay and all samples should be read off the standard curve. The end user should evaluate recoveries of corticosterone in other samples being tested. Moderate to severely hemolyzed samples should not be used in this kit.Ĭorticosterone can be assayed in other sample types by using one of the extraction protocols available on our website at: Corticosterone is identical across all species and we expect this kit may measure corticosterone from sources other than human. Samples containing visible particulate should be centrifuged prior to using. It has also been validated for dried fecal extract samples. This assay has been validated for serum, EDTA and heparin plasma, urine samples and for tissue culture samples. Software for converting raw relative optical density readings from the plate reader and carrying out four parameter logistic curve (4PLC) fitting. Repeater pipet with disposable tips capable of dispensing 25 μL, 50 μL and 100 μL.Ĭolorimetric 96 well microplate reader capable of reading optical density at 450 nm. Stop Solution A 1M solution of hydrochloric acid. Wash Buffer Concentrate A 20X concentrate that should be diluted with deionized or distilled water. 28 mL or 55 mLĭissociation Reagent is to be used only with Serum and plasma samples. 3 mL or 13 mLĪssay Buffer Concentrate A 5X concentrate that must be diluted with deionized or distilled water. 3 mL or 13 mLĭetectX® Corticosterone Conjugate A corticosterone-peroxidase conjugate in a special stabilizing solution.
125 or 625 μLĭetectX® Corticosterone Antibody A sheep polyclonal antibody specific for corticosterone. 1 or 5 EachĬorticosterone Standard Corticosterone at 100,000 pg/mL in a special stabilizing solution. In addition to stress levels, corticosterone is believed to play a decisive role in sleep-wake patterns.Ĭoated Clear 96 Well plates A clear plastic microtiter plate(s) coated with donkey anti-sheep IgG. Studies involving corticosterone and levels of stress include impairment of long term memory retrieval, chronic corticosterone elevation due to dietary restrictions and in response to burn injuries. Corticosterone is a major indicator of stress and is the major stress steroid produced in non-human mammals. Corticosterone is produced in response to stimulation of the adrenal cortex by ACTH and is the precursor of aldosterone. Corticosterone is a glucocorticoid secreted by the cortex of the adrenal gland. After a short incubation, the reaction is read at 450nm.
The substrate reacts with the bound corticosterone-peroxidase conjugate. After an hour incubation the plate is washed and substrate is added. The binding reaction is initiated by the addition of a polyclonal antibody to corticosterone. A corticosterone-peroxidase conjugate is added to the wells. Diluted samples are pipetted into a clear microtiter plate coated with an antibody. The kit has been developed for extracted fecal samples and has been validated with multiple animal species. The Corticosterone Enzyme Immunoassay kit measures corticosterone present in tissue culture, serum, plasma and fecal samples. Sample Types validated: Dried Fecal Extracts, Serum, EDTA and Heparin Plasma and Tissue Culture Media Serum, Plasma (EDTA), Plasma (heparin), Urine, Fecal, Tissue Culture Medium The DetectX® Corticosterone Immunoassay kit is designed to quantitatively measure Corticosterone present in extracted dried fecal samples, serum, plasma and tissue culture media samples. Annexin V-FITC Apoptosis Detection Kits.
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