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Corticosterone EIA Kit

Manufactured by Arbor Assays
Sourced in United States

The Corticosterone EIA Kit is a laboratory equipment product that provides a quantitative method for measuring corticosterone levels in biological samples. The kit utilizes an enzyme-linked immunosorbent assay (EIA) technique to detect and quantify corticosterone concentrations.

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12 protocols using Corticosterone EIA Kit

Once restrained, tail blood was collected from each rat at 0 min, 30 min into, and at the end (3 h) of the acute traumatic stressor. Specifically, a sterile scalpel was used to remove a small segment at the end of the tail, and approximately 0.1–0.2 mL of blood was collected at each time point. For the baseline time point at the start of the stressor, we collected blood within the first 2 min to avoid detecting elevations in corticosterone due to the restraint itself. All collected samples were kept on ice throughout the stressor. Blood clots were then removed, and samples were centrifuged at 9391 g for 20 min at 4 °C. Serum was then extracted and stored in clean tubes at −80 °C. Samples were assayed using a Corticosterone EIA kit (Arbor Assays, Ann Arbor, MI), with 2 replicates per sample. Samples were compared against a standard curve to obtain the concentration of corticosterone within each sample. Using corticosterone values from the three time points throughout stress exposure (0, 30 and 180 min), we calculated an area under the curve (AUC) for each animal. This provides us with an overall measure of the corticosterone response.
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Whole blood was collected by either cardiac puncture or retro-orbital bleeding and placed into Micro tube Z gel (41.1378.005; Sarstedt). Serum was separated by centrifuge at 10,000g for five minutes and stored at −80 °C until use. Corticosterone levels were detected using the Corticosterone EIA Kit (K014-H5; Arbor Assays) according to the manufacturer’s protocol. Owing to the effects of the circadian rhythm on corticosterone levels, we collected blood samples only between 13:00 and 17:00 on each day of the experiment.
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ELISA for detecting IGF1, IL-6, IL-7, and total corticosterone was performed using the IGF-I mouse/rat ELISA kit (Mediagnost), the Mouse IL-6 Quantikine ELISA kit (R&D Systems), the Mouse IL-7 Quantikine ELISA kit (R&D Systems), and Corticosterone EIA kit (ARBOR ASSAYS), respectively, according to the manufacturer’s instructions.
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Mice were exposed to carbon dioxide (~2 min) until they stopped breathing. The blood was then collected at the decapitation site using EDTA-coated tubes. Plasma was stored at −80 °C until use. Hormones were measured using the corticosterone EIA kit (K014-H1, Arbor Assays, Ann Arbor, MI, USA) and the mouse ACTH ELISA kit (NBP3-14759, Novus Biologicals, Centennial, CO, USA) according to manufacturers’ instructions. Five (corticosterone) and 25 (ACTH) microliters of plasma from each mouse were measured for each data point.
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Serum corticosterone levels were measured using commercially available enzyme immunoassay (EIA) kits (Corticosterone EIA Kit, Arbor Assays, Ann Arbor, MI, USA). Serum adrenaline levels were determined using a commercial kit (Adrenaline Research ELISA, LDN, Nordhorn, Germany). Absorbances were measured at 450 and 570 nm using a UV spectrophotometer (Molecular Devices, Sunnyvale, CA, USA).
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3-hydroxybutyrate concentrations were quantified in serum using a Roche Cobas Fara II analyzer (Roche Diagnostic Systems; Montclair, NJ). Corticosterone concentrations were quantified in serum using Corticosterone EIA Kit (Arbor Assays Inc, MI).
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Serum FGF21 concentrations were determined using Rat/Mouse FGF21 enzyme-linked immunosorbent assay (ELISA) kits (#MF2100, R&D Systems, Minneapolis, MN, USA). Serum corticosterone concentrations were determined using a Corticosterone EIA Kit (#K014-H1, Arbor Assays, Ann Arbor, MI, USA).
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Whole submandibular (chin) blood of non-anesthetized mice was collected using lancets and Eppendorf tubes. Blood samples were collected between 9:00 am and 12:00 pm. Plasma was separated by centrifugation at 10,000 g at 4°C for five minutes and stored at −80 °C until assayed. Corticosterone levels were detected using the Corticosterone EIA Kit (K014-H5; Arbor Assays) according to the manufacturer’s protocol.
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Serum samples were collected at 12:00 h, before animals were killed and stored at −80°C until further examination. Blood was collected by cardiac puncture and approximately 0.8 mL blood was collected from each mouse. After collection, blood was transferred to autoclaved 1.5 mL microcentrifuge tubes, followed by centrifugation at 4°C (13,400 g, 20 min). The transparent supernatant was then transferred to a fresh microcentrifuge tube as serum. Levels of corticosterone and adrenocorticotropic hormone (ACTH) were determined with commercial enzyme immunoassay kits (Mouse ACTH ELISA, Sigma Aldrich; Corticosterone EIA Kit, Arbor Assays, Ann Arbor, MI, USA).
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At 0 min, 30 min, and 3 h into acute immobilization stress, tail vein blood was collected from each rat for corticosterone sampling. Blood samples were centrifuged at 9391g for 20 min at 4 °C, and serum was extracted and stored at −80 °C. Samples were assayed using a Corticosterone EIA kit (Arbor Assays, Ann Arbor, MI). Any sample running below the detection limit of the assay was assigned the value of the detection limit (15.625 ng/mL).
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