After 12 h of food deprivation, rat serum was obtained to analyze insulin (ELISA, Millipore, Billerica, MA, USA), GLP-1 (ELISA, Millipore, Billerica, MA, USA), total cholesterol (TC, enzyme end-point method; Roche Diagnostics, GmbH, Mannheim, Germany), triglyceride (TG, enzyme end-point method, Roche Diagnostics, GmbH, Mannheim, Germany), and IL-6 (ELISA, Millipore, Billerica, MA, USA) concentrations, according to the manufacturer’s instruction. The homeostasis model assessment of insulin resistance (HOMA-IR) was calculated by the following formula: fasting blood glucose (FBG, mmol/L) × fasting serum insulin (µIU/mL)/ 22.5.
Elisa
Manufactured by Merck Group
Sourced in United States, Germany, United Kingdom, Sao Tome and Principe
ELISA is an analytical biochemistry assay that uses antibodies to detect and quantify the presence of a substance, typically a protein, in a liquid sample or wet sample. The core function of ELISA is to measure the concentration of a target analyte in a sample.
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Lab products found in correlation
Elisa, by R&D Systems (23 mentions)
Elisa, by Mercodia (7 mentions)
Ysi 2300, by Yellow Springs Instruments (5 mentions)
Protease inhibitor cocktail, by Merck Group (5 mentions)
Dpp4 inhibitor, by Merck Group (5 mentions)
Radioimmunoassay, by Merck Group (5 mentions)
Colorimetric assay, by Fujifilm (4 mentions)
Elisa, by ALPCO (4 mentions)
Onetouch ultraeasy, by LifeScan (4 mentions)
Ria assay, by Merck Group (4 mentions)
Glucocard glucometer, by Arkray (4 mentions)
Luminex, by Merck Group (3 mentions)
Solid phase sandwich elisa, by R&D Systems (3 mentions)
Accu check, by Roche (3 mentions)
Onetouch ultra2 glucometer, by LifeScan (3 mentions)
Colorimetric assay, by Roche (3 mentions)
Elitech assay kit, by ELITechGroup (3 mentions)
Accu chek, by Roche (3 mentions)
Elisa kit, by Merck Group (3 mentions)
Aprotinin, by Merck Group (3 mentions)
399 protocols using elisa
1
Serum Metabolite Analysis in Fasted Rats
2
Plasma Biomarker Quantification Protocol
3
Biomarker Analysis of Blood Samples
4
Plasma Biomarker Measurement Protocol
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Blood was collected in chilled tubes containing EDTA (7.4 mmol/L; glucose, insulin and C‐peptide) or EDTA and the DPP‐4 inhibitor diprotin A (0.1 mmol/L; Bachem, Bubendorf, Switzerland; GIP and GLP‐1). Samples were immediately centrifuged at 4°C, and plasma was frozen at −20°C. Glucose was measured using the glucose oxidase method, insulin using ELISA (Mercodia, Uppsala, Sweden, detection limit 6.9 pmol/L, interassay coefficient of variation [CV] 2.6%‐3.6% and intraassay CV 2.1%‐2.9% at low and high levels), total GIP using ELISA (Merck, detection limit 0.8 pmol/L, interassay CV 3.0%‐8.8% and intraassay CV 2.1%‐3.9% at low and high levels), and total GLP‐1 using ELISA (Merck, Burlington, MA, USA, detection limit 1.5 pmol/L, interassay CV 2.8%‐3.2% and intra‐assay CV 1.0%‐2.0%). Paracetamol was analysed colorimetrically (Cambridge Life Science, Ely, Cambridgeshire, UK).
5
Measurement of Ghrelin and IGF-1 Levels
6
Assay of Appetite Hormones in Plasma
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Blood samples were collected in the appropriate red or lavender topped tubes and Pefabloc tubes at our Institution’s TCRC and processed accordingly by the TCRC lab into 2 mL serum and plasma aliquots after centrifuging. For ghrelin assays, the plasma from the Pefabloc tubes was centrifuged and then transferred into HCL tubes before being aliquoted into cryovials. The aliquots were stored in −80 °C freezers and tested in a single batch for assay of appetite hormones. Serum aliquots were used to assay PYY, and plasma tubes with HCL additive were used for the ghrelin assay. Serum ghrelin concentrations were measured by an enzyme-linked immunosorbent assay (ELISA) (EMD Millipore Corporation, MO, USA; intra-assay coefficient of variation (CV) of 1.32%, inter-assay CV of 6.62%, and sensitivity of 50 pg/mL). PYY serum concentrations were also measured by an ELISA (Millipore Corporation (Linco Research), Billierca, MA, USA; intra-assay CV of 17–18%, inter-assay CV of 12–18%, and sensitivity of 10 pg/mL).
7
Metabolic Profiling and Growth Hormone Response
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Measurement of body weights and tail vein blood collection were performed as previously described [49] (link). Levels of serum leptin were determined by ELISA (Millipore). Fat mass was quantified using an EchoMRI Quantitative Whole Body Composition analyzer (Zinsser Analytic) on unanesthetized animals. Assessment of food intake and leptin sensitivity was performed in singly housed mice for 3 consecutive experimental days as previously described [49] (link) using 1.5 mg/kg recombinant mouse leptin (R&D Systems).
Metabolic rate was measured by indirect calorimetry with the use of a comprehensive lab animal monitoring system (CLAMS, Columbus Instruments). Mice were singly housed, with energy expenditure (using constants according to [50] (link)) determined over a 96-hour period. Core body temperature was assessed using a rectal thermometer (MM2000 digital thermometer, TME Thermometers).
For stimulation of growth hormone secretion (GH) in vivo, overnight fasted mice were given a single intraperitoneal injection of insulin (0.4 IU/kg) the following morning to induce hypoglycemia. Tail vein blood was collected prior to, and 1 h after, the insulin injection. GH levels in whole pituitary and in serum were assessed by ELISA (Millipore).
Metabolic rate was measured by indirect calorimetry with the use of a comprehensive lab animal monitoring system (CLAMS, Columbus Instruments). Mice were singly housed, with energy expenditure (using constants according to [50] (link)) determined over a 96-hour period. Core body temperature was assessed using a rectal thermometer (MM2000 digital thermometer, TME Thermometers).
For stimulation of growth hormone secretion (GH) in vivo, overnight fasted mice were given a single intraperitoneal injection of insulin (0.4 IU/kg) the following morning to induce hypoglycemia. Tail vein blood was collected prior to, and 1 h after, the insulin injection. GH levels in whole pituitary and in serum were assessed by ELISA (Millipore).
8
Metabolic and Stress Responses in LV-treated Rats
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To determine changes in metabolic parameters, LV-treated rats were subjected to an oral glucose tolerance test (OGTT) as described in our previous studies (12 (link),15 (link)). A cohort of rats was subjected to an overnight fast, and the following morning a blood sample was collected for baseline measures. Glucose (2 g/kg) was then administered by gastric intubation. Blood samples were collected from the tip of the tail at the following time points after intubation: 30, 60, 90, and 120 min. Blood glucose levels were measured by the glucose oxidase method (Pointe Scientific, Inc., Canton, MI); plasma insulin levels were measured by ELISA (Millipore, Billerica, MA).
To determine stress responses, a separate cohort of LV-treated rats were subjected to an acute restraint stress as described in our previous studies (16 (link)). Plasma corticosterone (CORT) levels were determined by ELISA (Enzo, Farmingdale, NY). Plasma leptin levels were also determined by ELISA (Millipore) in LV-Con and LV-IRAS rats (12 (link),15 (link)). ELISA plates were analyzed according to the manufacturer’s instructions using a Tecan SPECTRAFluor plate reader (Tecan U.S., Inc., Durham, NC). Statistical analysis was performed using a two-tailed, unpaired Student t test with P < 0.05 as the criterion for statistical significance.
To determine stress responses, a separate cohort of LV-treated rats were subjected to an acute restraint stress as described in our previous studies (16 (link)). Plasma corticosterone (CORT) levels were determined by ELISA (Enzo, Farmingdale, NY). Plasma leptin levels were also determined by ELISA (Millipore) in LV-Con and LV-IRAS rats (12 (link),15 (link)). ELISA plates were analyzed according to the manufacturer’s instructions using a Tecan SPECTRAFluor plate reader (Tecan U.S., Inc., Durham, NC). Statistical analysis was performed using a two-tailed, unpaired Student t test with P < 0.05 as the criterion for statistical significance.
9
Plasma Biomarker Analysis Protocol
10
Hepatic Lipid and Metabolic Biomarker Quantification
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Lipids were extracted from liver using a modified Folch method.30 (link) Hepatic triglyceride and cholesterol levels were quantified by colorimetric assay.17 (link)31 (link) Plasma insulin, leptin, and adiponectin concentration levels were determined at baseline, 8 weeks, and at sacrifice through enzyme-linked immunosorbent assay (ELISA; Millipore, Billerica, MA, USA) according to manufacturer's instructions.17 (link) Plasma interleukin-6 (IL-6) concentrations also were measured at study termination through a commercially available ELISA.
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