Corticosterone high sensitivity eia kit

Manufactured by Immunodiagnostic Systems

Sourced in Switzerland, United States, United Kingdom

The Corticosterone High Sensitivity EIA kit is a laboratory equipment product designed for the quantitative determination of corticosterone levels in various sample types. It utilizes an enzyme immunoassay (EIA) technique to measure corticosterone concentrations.

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Lab products found in correlation

Edta treated capillary tubes, by Sarstedt (1 mentions) Accu check aviva, by Roche (1 mentions) Corticosterone enzyme immunoassay kit, by Arbor Assays (1 mentions) Multiskan spectrum, by Thermo Fisher Scientific (1 mentions) Ultracentrifuge, by Beckman Coulter (1 mentions)

3 protocols using corticosterone high sensitivity eia kit

Restraint Stress Test and Corticosterone Assay

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On PND42 (± 1), we performed a restraint stress test by placing the animals in a transparent tube (according to the size of the animal: Ø2.86cm; BioSeb, Vitrolles, France or Ø2.48cm; G&P Kunsstofftechnik, Kassel, Germany) for 60 min. Blood (approx. 50µL) was collected (in EDTA treated capillary tubes; Sarstedt, Nümbrecht, Germany) by incision of a lateral tail vein, immediately after placing and approximately 1 min before releasing the animal from the restrainer. Plasma was subsequently separated by one-step centrifugation (2000 × g, 10 min, 4 °C) and stored at − 80 °C until analysis. Glucose measurements were performed immediately using a standard blood glucose meter (Accu-Check Aviva; Roche, Basel, Switzerland).
Corticosterone competitive ELISA.
Plasma corticosterone levels were measured using Corticosterone High Sensitivity EIA kit (Immunodiagnosticsystems) according to the manufacturer’s instructions. Samples were diluted in assay diluent, either 50-fold (baseline samples pre-stress) or 100-fold (one hour of restraint).

Merz M.P., Seal S.V., Grova N., Mériaux S., Guebels P., Kanli G., Mommaerts E., Nicot N., Kaoma T., Keunen O., Nazarov P.V, & Turner J.D. (2024). Early-life influenza A (H1N1) infection independently programs brain connectivity, HPA AXIS and tissue-specific gene expression profiles. Scientific Reports, 14, 5898.

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Corticosterone Quantification in Lizard Plasma

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Plasma samples from the two laboratory studies were analyzed for CORT concentrations using the Corticosterone High Sensitivity EIA Kit (Immunodiagnostic Systems Ltd., Fountain Hills, AZ, USA). This kit was previously validated in this species^{39 (link)}, and each sample was diluted 1:10 with assay buffer to ensure tested samples fell within the range of the standard curve. All samples were run in duplicate, with intra-assay coefficients of variation of 7.3–15.9 and an inter-assay coefficient of variation of 14.8.
Plasma samples from field-caught fence lizards were analyzed using the Corticosterone Enzyme Immunoassay Kit (Catalog number K014; Arbor Assays, Ann Arbor, MI, USA). Plasma was diluted 1:100 per manufacturer’s instructions, to ensure tested samples fell within the range of the standard curve. Percent recovery of CORT for this kit was approximately 95%, with good parallelism between the standard curve and a dilution curve made from pooled fence lizard plasma samples (CT unpubl. data). Each sample was run in duplicate, with intra-assay coefficients of variation of 5.6–9.2 and an inter-assay coefficient of variation of 6.3.

Racic A., Tylan C, & Langkilde T. (2020). Effects of temperature on plasma corticosterone in a native lizard. Scientific Reports, 10, 16315.

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Cytoplasmic Fraction Isolation and Corticosterone Determination

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For the preparation of cytoplasmic fraction from VAT, tissue was homogenized (w/v = 1:1) in ice-cold homogenization buffer (20 mM Tris-HCl, pH 7.2, 10% glycerol, 50 mM NaCl, 1mM EDTA-Na 2 , 1 mM EGTA-Na 2 , 2 mM DTT, protease and phosphatase inhibitors). The cell lysate was filtered through gauze and centrifuged at low speed (2000 g, 15 min, 4°C). The resulting supernatant was centrifuged (10,000 g, 25 min, 4°C), recentrifuged (150,000 g, 90 min, 4°C, Beckman ultracentrifuge) and the final supernatant was used as cytoplasmic fraction. Lowry method was used for the determination of protein concentration (Lowry et al. 1951) .
VAT corticosterone (CORT) was assessed using Corticosterone High Sensitivity EIA kit according to manufacturer's instructions (Immunodiagnostic Systems LTD, Tyne and Wear, UK). Absorbance at 450 nm (reference 650 nm) was measured spectrophotometrically (Multiskan Spectrum, Thermo, Finland). CORT concentrations were determined using four parameter logistic (4PL) curvefitting method (Prism, GraphPad Software) and given as ng/mg of protein. The assay sensitivity was 0.17 ng/mL, while intra-assay and inter-assay CVs were 5.9 and 8.9%, respectively.

Gligorovska L., Bursać B., Kovačević S., Veličković N., Matić G, & Djordjevic A. (2019). Mif deficiency promotes adiposity in fructose-fed mice. The Journal of endocrinology, 240(2).

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