The concentrations of galanin, LH, FSH, testosterone, SHBG, cortisol, and corticosterone in rat sera were detected using Rat Galanin ELISA Kit (#MBS2512765, Mybiosource, San Diego, USA), Rat LH ELISA Kit (#MBS2018978, Mybiosource), Rat FSH ELISA Kit (#EKU04249, Biomatik, Ontario, Canada), Testosterone ELISA Kit (#582701, Cayman Chemical, Michigan), Rat SHBG ELISA Kit (#MBS261678, Mybiosource), Cortisol Parameter Assay Kit (#KGE008B, R&D systems, Minnesota, USA), and Corticosterone Parameter Assay Kit (#KGE009, R&D), respectively. The concentrations of TG, total CHOL, HDL-C, LDL-C, and NEFA in rat sera and liver tissues were detected using Triglyceride Quantification Assay Kit (#ab65336, Abcam), HDL and LDL/VLDL Cholesterol Assay Kit (#ab65390, Abcam), Rat LDL-C ELISA Kit (#CSB-E16561r, Cusabio, Wuhan, China), and Free Fatty Acid Quantification Assay Kit (#ab65341, Abcam), respectively. We carried out all procedures following the recommended instructions provided by kit’s manufacturers.
Corticosterone parameter assay kit
Manufactured by R&D Systems
Sourced in United States
The Corticosterone Parameter Assay Kit is a lab equipment product that can be used to measure corticosterone levels in various sample types. The kit utilizes a competitive enzyme immunoassay (EIA) principle to quantify corticosterone concentration.
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Lab products found in correlation
Cortisol parameter assay kit, by R&D Systems (1 mentions)
Testosterone elisa kit, by Cayman Chemical (1 mentions)
Hdl and ldl vldl cholesterol assay kit, by Abcam (1 mentions)
Free fatty acid quantification assay kit, by Abcam (1 mentions)
Triglyceride quantification assay kit, by Abcam (1 mentions)
Mouse il 1 beta il 1f2 quantikine elisa kit, by R&D Systems (1 mentions)
Mouse il 6 quantikine elisa kit, by R&D Systems (1 mentions)
Quantichrom creatinine assay kit, by BioAssay Systems (1 mentions)
Duoset elisa, by R&D Systems (1 mentions)
Pierce chromogenic endotoxin quant kit, by Thermo Fisher Scientific (1 mentions)
9 protocols using corticosterone parameter assay kit
1
Comprehensive Rat Serum and Tissue Analysis
2
Quantifying Corticosterone in EAE Model
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Corticosterone was measured in the control (n = 6) and EAE animals (n = 6/group) in serum acquired from blood samples obtained by cardiac puncture of at least six animals per group. Blood was centrifuged at 3,000 × g for 15 min, and all samples were stored at −80°C before determining corticosterone concentration using the commercially available Corticosterone Parameter Assay Kit (R&D Systems Inc., Minneapolis, MN, United States). The sensitivity of this kit is 0.047 ng/ml. The plate was read at 450 nm and wavelength correction was set to 570 nm. To determine the corticosterone level, calculation was performed using a four-parameter logistic curve fitting program.
3
Corticosterone Assay in Mice
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Serum or urine was collected from mice for the corticosterone assay. For serum, whole blood samples were collected by cardiac puncture after carbon dioxide euthanasia and placed on ice for 30 min. After centrifugation at 2000 × g 4 °C for 10 min, the supernatant was collected and frozen before the corticosterone assay. For urine, mice were transferred to single cages without bedding. After 1 h, urine on the cage bottom was collected. The test was performed using the Corticosterone Parameter Assay Kit (KGE009, R&D Systems) according to the manufacturer's protocol.
4
Quantifying Blood Biomarkers in Mice
5
Urine Biomarker Analysis in Mice
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Body weights were measured and urine was collected before and after the IntelliCage adaptation period, at the end of the IntelliCage measurements (after the reversal training), and before and after LabMaster monitoring (Figure 5 ). The urine samples were collected at 7 am with a noninvasive method, in which the mouse was placed in an empty clean cage and the urine samples were collected by pipette immediately and stored in −70 °C until further analysis. Urine corticosterone levels were analyzed with ELISA kits according to the manufacturer’s instructions (Corticosterone Parameter Assay Kit, R&D Systems®, Minneapolis, MN, USA). Urine creatinine levels were measured with QuantiChrom™ Creatinine Assay kit (BioAssay Systems, Hayward, California, United States) according to the manufacturer protocol. Urine creatinine levels were used to correct the urine corticosterone concentration for differences in kidney function such as glomerular filtration and hydration. The mice were euthanized by cervical dislocation after the PhenoMaster study.
6
Measuring Diurnal Leptin and Corticosterone
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Since daily changes in serum levels of leptin and corticosterone have been reported previously (17,18), the blood samples in this study were collected between 9:00 AM and 11:00 AM each day to reduce the occurrence of possible errors. Serum leptin and corticosterone levels were measured using the EzWay Mouse Leptin enzyme-linked immunosorbent assay (ELISA) kit (Cosmo bio, San Diego, CA, USA) and the Corticosterone Parameter Assay Kit (R&D Systems, Minneapolis, MN, USA) according to the manufacturers' instructions.
7
Corticosterone Quantification Protocol
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Serum corticosterone was determined using the Corticosterone Parameter Assay kit (R&D, KGE009) following the manufacturer’s instructions.
8
Quantification of Endotoxin, Inflammatory Markers, and Corticosterone in Murine Tissues
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Liver LPS levels were enumerated using a Pierce™ Chromogenic Endotoxin Quant Kit (Thermo Fisher Scientific, MD, USA). Values are represented as endotoxin units (EU). Mouse Lcn2/NGAL and S100a8 in tissue homogenates were enumerated using a DuoSet ELISA (R&D Systems, MN, USA) as per the manufacturer’s protocol. Numerical values were determined using an 8-point standard curve of known concentrations. Mouse IL-22 in ileal explant supernatant was enumerated using an ELISA (ThermoFisher) as per the manufacturer’s protocol. Numerical values were calculated using an 8-point standard curve of known concentrations. Mouse corticosterone in the serum was enumerated using a Corticosterone Parameter Assay Kit (R&D Systems) as per the manufacturer’s protocol. Numerical values were determined using a 6-point standard curve of known concentrations, using an EnVision 2104 Multilabel Reader (v1.13). Serum samples were collected after 16 h of stress or overnight starvation and analyzed at Eve Technologies (Alberta, Canada) for cytokine and chemokine determinations using multiplex analyses. Samples are represented using a mean z-score.
9
Chronic Restraint Stress in Mice
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Mice were placed in 50 mL conical tubes and restrained at least 16 hr and suffered by heating at 42 C for 30 min for more than four consecutive days. Control mice were left in the home cages undisturbedly without food and water at the same time. All murine blood samples for determination of glucocorticoid in plasma were collected following the stress, and the corticosterone parameter assay kit (R&D systems, USA) was used for the measurement according to manufacturer's instructions. For open field test (OFT), mice were placed in an open field of a 45 3 45 3 45 cm box and analyzed for activities. Mice were monitored at least 10 min and recorded the amount of time they spent in the center or peripheral regions of the box. For novelty suppressed feeding test (NSFT), mice were placed in one of the corners of the box after 24-hr fasting. In the maximum of 10 min, the latency to the first bite of chow placed in the middle of the box was recorded. Then the box was cleaned with 70% alcohol prior to testing, and fresh food was used for each mouse.
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