Blood collected from mice injected with DB or irisin twice a week for 4 weeks were incubated at room temperature (RT) for 30 min. Blood samples were then centrifuged at 1500×g for 15 min at 4 °C and serum was isolated. Irisin levels in the serum were quantified using a commercial ELISA kit following the manufacturer’s protocol (Irisin ELISA kit EK-067−52, Phoenix Pharmaceuticals, Inc., USA) using spectrophotometry at a wavelength of 450 nm. The detection range of the kit is 0.1–1000ng/ml.
Irisin elisa kit ek 067 52
Manufactured by Phoenix Pharmaceuticals
Sourced in United States
The Irisin ELISA Kit EK-067-52 is a quantitative sandwich enzyme-linked immunosorbent assay (ELISA) designed for the detection and quantification of irisin levels in biological samples. The kit utilizes a specific antibody coated on a 96-well plate to capture irisin, which is then detected using a detection antibody conjugated with an enzyme. The enzymatic reaction produces a color change proportional to the amount of irisin present in the sample.
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Lab products found in correlation
Immunoradiometric assay, by Thermo Fisher Scientific (2 mentions)
Hexokinase activity assay, by Abcam (1 mentions)
Triglycerides glycerol blanked, by Roche (1 mentions)
Dimension autoanalyzer, by Roche (1 mentions)
Ldl cholesterol plus 2nd generation, by Roche (1 mentions)
Cholesterol gen 2, by Roche (1 mentions)
4 protocols using irisin elisa kit ek 067 52
1
Quantification of Serum Irisin Levels
2
Quantifying Irisin in Human Plasma
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Blood samples from overall participants were obtained under basal conditions after a 12-h overnight fast. EDTA-plasma and serum of specimen were separated from whole blood and immediately frozen at −80° C until assay.
The quantitative measurement of irisin in human plasma samples was performed using a commercial enzyme-linked immunosorbent assay (ELISA) kit directed against amino acids 31–143 of the FNDC5 protein (Irisin ELISA Kit EK-067-52; Phoenix Pharmaceuticals, INC, CA) according to the manufacturer’s instructions. Absorbance from each sample was measured in duplicate using a spectrophotometric microplate reader at wavelength of 450 nm (Versamax Microplate Reader; Associates of Cape Cod Incorporated, East Falmouth, MA).
The quantitative measurement of irisin in human plasma samples was performed using a commercial enzyme-linked immunosorbent assay (ELISA) kit directed against amino acids 31–143 of the FNDC5 protein (Irisin ELISA Kit EK-067-52; Phoenix Pharmaceuticals, INC, CA) according to the manufacturer’s instructions. Absorbance from each sample was measured in duplicate using a spectrophotometric microplate reader at wavelength of 450 nm (Versamax Microplate Reader; Associates of Cape Cod Incorporated, East Falmouth, MA).
3
Quantitative Irisin Measurement in Plasma
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The quantitative measurement of irisin in human plasma samples was performed using a commercial enzyme-linked immunosorbent assay (ELISA) kit directed against amino acids 31–143 of the FNDC5 protein (Irisin ELISA Kit EK-067-52; Phoenix Pharmaceuticals Inc., CA) according to the manufacturer's instructions. The absorbance from each sample was measured in duplicate using a spectrophotometric microplate reader at wavelength of 450 nm (Versamax Microplate Reader; Associates of Cape Cod Incorporated, East Falmouth, MA).
Insulin was analysed by an immunoradiometric assay (BioSource International, Camarillo, CA, USA) in a Beckman Coulter (Fullerton, CA, USA), showing 0.3% cross-reaction with proinsulin. The intra- and interassay CV were 1.9% and 6.3%, respectively. Glucose was measured using a Dimension Autoanalyzer (Dade Behrng, Deerfield, IL, USA). The homeostatic model assessment index (HOMA-IR) was calculated following the formula (fasting plasma glucose (mg/mL) × fasting plasma insulin (mU/L)/405), as described elsewhere [27 (link)].
Insulin was analysed by an immunoradiometric assay (BioSource International, Camarillo, CA, USA) in a Beckman Coulter (Fullerton, CA, USA), showing 0.3% cross-reaction with proinsulin. The intra- and interassay CV were 1.9% and 6.3%, respectively. Glucose was measured using a Dimension Autoanalyzer (Dade Behrng, Deerfield, IL, USA). The homeostatic model assessment index (HOMA-IR) was calculated following the formula (fasting plasma glucose (mg/mL) × fasting plasma insulin (mU/L)/405), as described elsewhere [27 (link)].
4
Serum Irisin and Metabolic Markers
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Participants underwent blood sampling for the assessment of blood chemistry and hormonal parameters. The quantitative measurement of irisin in human serum samples was performed using a commercial enzyme linked immunosorbent assay (ELISA) kit (Irisin ELISA Kit EK-067-52; Phoenix Pharmaceuticals Inc., CA) according to the manufacturer’s instructions. Basal insulin was analyzed by an immunoradiometric assay (BioSource International, Camarillo, CA, USA) in a Beckman Coulter (Fullerton, CA, USA). Enzymatic colorimetric assay was used to measure fasting plasma glucose (FPG) (Hexokinase activity assay, Abcam), total cholesterol (Cholesterol Gen.2, Roche), high-density lipoproteins (HDL) cholesterol (LDL-Cholesterol plus 2nd generation, Roche), triglycerides (Triglycerides/Glycerol Blanked, Roche) concentrations using a Dimension Autoanalyzer (Cobas501, Roche, Switzerland). The homeostatic model assessment index was calculated following the formulas: HOMA-IR = fasting serum insulin (FINS, mU/L) × fasting plasma glucose (FPG, mmol/L)/22.5; HOMA-IS = 1/(FPG (mmol/L) × FINS (mIU/L)). Low-density lipoprotein (LDL) cholesterol serum concentration was calculated with Friedewald’s formula.
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