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Corticosterone elisa kit

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The Corticosterone ELISA Kit is a quantitative in-vitro diagnostic test used to measure the concentration of corticosterone in biological samples. The kit utilizes the competitive enzyme-linked immunosorbent assay (ELISA) principle to determine the corticosterone levels.

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Lab products found in correlation

Vacutainer, by BD (4 mentions) Synergy ht, by Agilent Technologies (3 mentions) Phospholipids c test, by Fujifilm (2 mentions) Cholesterol c test, by Fujifilm (2 mentions) Glucose cii test, by Fujifilm (2 mentions) Multi array mouse rat insulin and leptin kit, by Mesoscale (1 mentions) Cobas mira autoanalyzer, by Roche (1 mentions) Nefa c test wako, by Fujifilm (1 mentions) Triglyceride e test wako, by Fujifilm (1 mentions) Cholesterol e test wako, by Fujifilm (1 mentions) Glucocard g meter, by Arkray (1 mentions) Tba test, by Fujifilm (1 mentions) Triglyceride e test, by Fujifilm (1 mentions) Acth elisa kit, by Phoenix Pharmaceuticals (1 mentions) Insulin elisa kit, by Mercodia (1 mentions) Synergy ht multi mode microplate reader, by Agilent Technologies (1 mentions) Ethanol assay kit, by BioAssay Systems (1 mentions)

13 protocols using corticosterone elisa kit

1

Validating Corticosterone Levels in Stress Model

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The blood plasma corticosterone level was analyzed to verify the validity of our model. Both the control and stress groups (Control, n = 10; 3-week stress, n = 10) were briefly anesthetized with 3% isoflurane immediately after termination of the last stress, and the cardiac blood was collected in heparin-coated tubes (BD Vacutainer, Becton Dickinson, NJ, USA). The blood samples were centrifuged at 3500 rpm for 15 min. The segregated plasma was diluted 1:200 and analyzed using a corticosterone ELISA kit (Assaypro LLC, MO, USA). The fluorescence intensity was scanned at 450 nm by a microplate luminescence reader (Synergy HT, BioTek, VT, USA). A corticosterone standard curve was generated using standard solutions, and then, the sample concentration was determined from the standard curve.
Lee S., Kang B.M., Shin M.K., Min J., Heo C., Lee Y., Baeg E, & Suh M. (2015). Chronic Stress Decreases Cerebrovascular Responses During Rat Hindlimb Electrical Stimulation. Frontiers in Neuroscience, 9, 462.
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2

Quantification of Stress Hormones and Metabolic Markers

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Corticosterone was measured using a commercially available corticosterone ELISA kit (AssayPro, St. Charles, MO, USA) according to the manufacturer's protocol. Glucose levels were analyzed by Cobas MIRA autoanalyzer (Hoffman LaRoche, Basel, Switzerland). Insulin and leptin levels were measured by using a Multi-Array Mouse/Rat Insulin and Leptin kit (Meso Scale Discovery, Gaithersburg, USA).
Lee S.J., Diener K., Kaufman S., Krieger J.P., Pettersen K.G., Jejelava N., Arnold M., Watts A.G, & Langhans W. (2016). Limiting glucocorticoid secretion increases the anorexigenic property of Exendin-4. Molecular Metabolism, 5(7), 552-565.
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3

Chick Plasma Biomarkers and Tissues

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At the end of the experiment and under carbon
dioxide anesthesia, blood was collected from the
carotid arteries of chicks and chicks were
euthanized by decapitation. Plasma was separated
immediately by centrifugation at 1,910 × g for 10
min at 4 °C and then stored at −80 °C until
analysis. Plasma corticosterone and uric acid
concentrations were measured using commercial kits
(Corticosterone ELISA Kit, AssayPro LLC, MO, USA;
Uric Acid C-Test, Code: 437-17301, Fujifilm Wako
Pure Chemicals Co., Osaka, Japan). The weights of
the breasts, legs, and livers were recorded. Then,
the center pieces of the PM, BF, and liver were
excised and immediately frozen in liquid nitrogen
and stored at −80 °C for real-time polymerase
chain reaction (PCR) analyses.
El-far A.S., Kamiya M., Saneyasu T, & Honda K. (2024). Effects of Amino Acid Supplementation to a Low-Protein Diet on the Growth Performance and Protein Metabolism-related Factors in Broiler Chicks. The Journal of Poultry Science, 61, 2024014.
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4

Corticosterone Measurement in RLED Exposed Mice

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Animals received 1ml/kg 10% Heparin, IP, 10 minutes before the RLED exposure ended. Upon completion of each animal’s RLED exposure each animal was taken to a laminar flow hood and induced on 4% Isoflurane in 2% O2 and maintained on 2% Isoflurane in 2% O2 during the tail vein blood collection. Warm water was applied to the tail and 300–400 μl of tail vein blood was collected and put into 1ml BD Microtainer® Green Topped Serum Separator Tubes with Lithium Heparin. Tail vein blood was collected within 5–7 minutes after the completion of the RLED exposure. Tubes were then spun down at 4 degrees Celsius at 3000 revolutions per minute for 10 minutes. Serum was transferred into microcentrifuge tubes on dry ice and transported to a −80 freezer and stored there until the time of the ELISA. Pre- and post-RLED exposure tail vein blood was collected between the hours of 9 am and 10 am for each of the animals. Animals were taken to a different room than the RLED exposure for the blood collection in the laminar flow hood and any blood was cleaned up from the hood surfaces between animals. The corticosterone ELISA kit was purchased from Assaypro, catalog #ABIN577656. The conjugate was stored separate from the rest of the kit at −20 degrees Celsius until the day the ELISA kit was run. All other components of the ELISA kit were stored at 4 degrees Celsius until the kit was run.
Khanna R., Patwardhan A., Yang X., Li W., Cai S., Ji Y., Chew L.A., Dorame A., Bellampalli S.S., Schmoll R.W., Gordon J., Moutal A., Vanderah T.W., Porreca F, & Ibrahim M.M. (2019). Development and Characterization of An Injury-free Model of Functional Pain in Rats by Exposure to Red Light. The journal of pain : official journal of the American Pain Society, 20(11), 1293-1306.
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5

Serum Metabolic and Hormonal Profiles

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Serum levels of glucose, lipids, and hormones were measured using appropriate equipment, reagents, and kits. The GLUCOCARD G+ meter was used to measure glucose content (Arkray, Kyoto, Japan). The NEFA C-Test Wako (Wako Pure Chemical Industries, Osaka, Japan) was used to measure free fatty acid levels. The Triglyceride E-Test Wako (Wako Pure Chemical Industries) was used to measure triglyceride levels and the Cholesterol E-Test Wako (Wako Pure Chemical Industries) for cholesterol content. The Rebis Insulin-rat T ELISA kit (Shibayagi, Gunma, Japan) was used to measure insulin levels, the Leptin ELISA kit (Morinaga Institute of Biological Science, Yokohama, Japan) for leptin, and the Corticosterone ELISA kit (Assaypro, St. Charles, MO) for corticosterone. Blood chemistry is listed in Tables 1 and 2.
Iwakoshi-Ukena E., Shikano K., Kondo K., Taniuchi S., Furumitsu M., Ochi Y., Sasaki T., Okamoto S., Bentley G.E., Kriegsfeld L.J., Minokoshi Y, & Ukena K. (2017). Neurosecretory protein GL stimulates food intake, de novo lipogenesis, and onset of obesity. eLife, 6, e28527.
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6

Liver Lipid Extraction and Analysis

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Liver lipids were extracted by using the method described by Folch et al. (36 (link)). Total liver lipids were measured gravimetrically. Hepatic cholesterol, triglycerides, and phospholipids in the lipid extract were measured enzymatically (T-CHO and TG-EN; Kainos Laboratories, Tokyo, Japan, Phospholipids C-Test, Wako Pure Chemical Industries, Osaka, Japan). Serum glucose, total cholesterol, triglyceride, non-esterified fatty acids (NEFA), and bile acids were measured enzymatically by using commercial kits (Glucose CII-test, Triglyceride E-test, Cholesterol C-test, NEFA C-test, TBA-test; Wako Pure Chemical Industries, Osaka, Japan). The insulin and corticosterone levels were measured using rat-specific enzyme immunoassay kits [Rat insulin ELISA kit (#M1101); Morinaga Institute of Biological Science, Yokohama, Japan, Corticosterone ELISA kit (#EC3001-1); Assaypro, MO, USA].
Kim D., Hanzawa F., Sun S., Laurent T., Ikeda S., Umeki M., Mochizuki S, & Oda H. (2021). Delayed Meal Timing, a Breakfast Skipping Model, Increased Hepatic Lipid Accumulation and Adipose Tissue Weight by Disintegrating Circadian Oscillation in Rats Fed a High-Cholesterol Diet. Frontiers in Nutrition, 8, 681436.
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7

Serum Biomarker Quantification by ELISA

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Levels of specific antibodies, corticosterone, ACTH, and insulin were measured in serum samples by an enzyme-linked immunosorbent assay (ELISA). We used a corticosterone ELISA kit (ASSAYPRO, St. Charles, MO), ACTH ELISA kit (Phoenix Pharmaceuticals, Burlingame, California), and an insulin ELISA kit (Mercodia, Uppsala, Sweden). Assays were performed according to the manufacturers’ instructions.
Sasaki H., Hattori Y., Ikeda Y., Kamagata M., Iwami S., Yasuda S., Tahara Y, & Shibata S. (2016). Forced rather than voluntary exercise entrains peripheral clocks via a corticosterone/noradrenaline increase in PER2::LUC mice. Scientific Reports, 6, 27607.
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8

Plasma Corticosterone Quantification

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After the 3-week RS paradigm, mice were briefly anesthetized with 3 % isoflurane, and approximately 200 µl of blood was collected in heparin-coated tubes (BD Vacutainer, Becton Dickinson, NJ). The blood samples were centrifuged at 13,000 rpm for 15 min at 4 °C. The concentration of corticosterone in plasma was analyzed using a corticosterone ELISA kit (Assaypro LLC, MO). The absorbance at 450 nm was measured using a microplate reader (Synergy HT Multi-Mode Microplate Reader, BioTek Instruments, Inc., VT). A standard curve was generated using standard solutions, and the plasma corticosterone level was determined from the standard curve.
Lee S., Kang B.M., Kim J.H., Min J., Kim H.S., Ryu H., Park H., Bae S., Oh D., Choi M, & Suh M. (2018). Real-time in vivo two-photon imaging study reveals decreased cerebro-vascular volume and increased blood-brain barrier permeability in chronically stressed mice. Scientific Reports, 8, 13064.
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9

Measuring Plasma Corticosterone in Mice

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After the end of 3-weeks of stress induction, plasma was collected without stress exposure on the day of collection. Mice (control, n = 25; stress, n = 25) were briefly anesthetized with 3% isoflurane (Hana Pharm) using an anesthesia machine (VetEquip), and trunk blood was collected in heparin-coated tubes (BD Vacutainer, Becton Dickinson). Blood samples were centrifuged at 5000 rpm for 10 min. The concentration of corticosterone in the plasma was analyzed using a corticosterone ELISA kit (Assaypro). Assay procedures were followed according to the instructions provided in the kit. The absorbance at a wavelength of 450 nm was scanned by a microplate reader (Synergy HT, BioTek). A standard curve was generated using standard solutions, and the sample concentrations were determined from the standard curve.
Han K., Min J., Lee M., Kang B.M., Park T., Hahn J., Yei J., Lee J., Woo J., Lee C.J., Kim S.G, & Suh M. (2019). Neurovascular Coupling under Chronic Stress Is Modified by Altered GABAergic Interneuron Activity. The Journal of Neuroscience, 39(50), 10081-10095.
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10

Quantifying Corticosterone Levels in Stressed Mice

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To quantify the plasma concentration of corticosterone, mouse plasma was collected immediately after the 30 min restraint stress period. Mice (control: n = 10; stress: n = 10) were anesthetized with Zoletil (30 mg/kg, i.p.), and heart blood was collected in heparin-coated tubes (BD Vacutainer, Becton Dickinson). The blood samples were centrifuged at 5000 rpm for 2 × 10 min at 4°C. The concentration of corticosterone in the plasma was measured using a corticosterone ELISA kit (Assaypro) according to the instructions provided in the kit. The absorbance was scanned at a wavelength of 450 nm using a microplate reader (Synergy HT, BioTek). A standard curve was generated using standard solutions, and the sample concentrations were calculated from the standard curve.
Han K., Lee M., Lim H.K., Jang M.W., Kwon J., Lee C.J., Kim S.G, & Suh M. (2020). Excitation-Inhibition Imbalance Leads to Alteration of Neuronal Coherence and Neurovascular Coupling under Acute Stress. The Journal of Neuroscience, 40(47), 9148-9162.
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