Mouse FGF-21 ELISA Kit
FGF21 is a metabolic hormone mainly produced in liver. It regulates glucose and lipid metabolism through pleiotropic actions in multiple tissues, and protects against metabolic damages under various stresses.
In humans, high circulating FGF21 levels are found in obesity and multiple cardiometabolic disorders, including metabolic syndrome, type 2 diabetes, non-alcoholic fatty liver disease and coronary artery disease. Serum FGF21 is a potential biomarker for the early detection and risk prediction of these diseases.
PRINCIPLE OF THE ASSAY
This assay is a quantitative sandwich ELISA. The immunoplate is precoated with a rabbit polyclonal antibody specific for mouse FGF-21. Standards and samples are pipetted into the wells and any mouse FGF-21 present is bound by the immobilized antibody. After washing away any unbound substances, a biotin labelled polyclonal antibody specific for mouse FGF-21 is added to the wells. After wash step to remove any unbound reagents, streptavidin-HRP conjugate (STP-HRP) is added. After the last wash step, an HRP substrate solution is added and colour develops in proportion to the amount of mouse FGF-21 bound initially. The assay is stopped and the optical density of the wells determined using a microplate reader. Since the increases in absorbance are directly proportional to the amount of captured mouse FGF-21, the unknown sample concentration can be interpolated from a reference curve included in each assay.
A. Typical representation of standard curve
The following standard curve is provided for demonstration only. A standard curve should be generated for each set of sample assay.
Absorbance (450 nm)
The lowest level of mouse FGF-21 that can be detected by this assay is 31.2 pg/mL.
Cross reactivity of recombinant proteins
Analyte Cross Reactivity Human FGF-21 No Mouse Mup-1 No Mouse LCN2 No Mouse Adiponectin No Mouse ANGPL4 No
Intra-assay Precision (Precision within an assay) C.V. <8%.
Inter-assay Precision (Precision between assays) C.V. <10%.
The recovery of the assay was determined by adding various amounts FGF21 to a sample. The measured concentration of the spiked sample in the assay was compared to the expected concentration. The average recovery was 92%.
To assess the linearity of the assay, a sample with high level of FGF-21 was serially diluted with the 1×Assay buffer to produce samples with values within the dynamic range of the assay.
Dilution Measured (pg/mL) Expected (pg/mL) Recovery (%) 1/2 1360 1360 100 1/4 680 680 101 1/8 380 340 111 1/16 200 170 115
PUBLICATIONS CITING THIS PRODUCT
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- Li H, Wu G, Fang Q, Zhang M, Hui X, Sheng B, Wu L, Bao Y, Li P, Xu A, Jia W. Fibroblast growth factor 21 increases insulin sensitivity through specific expansion of subcutaneous fat. Nature communications. 2018 Jan 18;9(1):1-6.
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- Geng L, Liao B, Jin L, Huang Z, Triggle CR, Ding H, Zhang J, Huang Y, Lin Z, Xu A. Exercise alleviates obesity-induced metabolic dysfunction via enhancing FGF21 sensitivity in adipose tissues. Cell reports. 2019 Mar 5;26(10):2738-52.
- Yang BC, Wu SY, Leung PS. Alcohol ingestion induces pancreatic islet dysfunction and apoptosis via mediation of FGF21 resistance. Annals of translational medicine. 2020 Mar;8(6).
- Zhang Y, Cao X, Chen L, Qin Y, Xu Y, Tian Y, Chen L. Exposure of female mice to perfluorooctanoic acid suppresses hypothalamic kisspeptin‐reproductive endocrine system through enhanced hepatic fibroblast growth factor 21 synthesis, leading to ovulation failure and prolonged dioestrus. Journal of neuroendocrinology. 2020 May;32(5):e12848.
- Zhou Y, Li C, Wang X, Deng P, He W, Zheng H, Zhao L, Gao H. Integration of FGF21 Signaling and Metabolomics in High-Fat Diet-Induced Obesity. Journal of Proteome Research. 2021 Jul 9;20(8):3900-12.
- Liu D, Gu J, Shao W, Pang J, Qian X, Jin T. Comparison of metabolic beneficial effects of Liraglutide and Semaglutide in male C57BL/6J mice. Canadian Journal of Diabetes. 2021 Sep 8.
- Wang Y, Zhang Y, Shi Z, Di T, Yu W, Chen L. Exposure of male mice to perfluorooctanoic acid induces anxiety-like behaviors by increasing corticotropin-releasing factor in the basolateral amygdala complex. Chemosphere. 2022 Jan 1;287:132170
- Badakhshi Y, Shao W, Liu D, Tian L, Pang J, Gu J, Hu J, Jin T. Estrogen-Wnt signaling cascade regulates expression of hepatic fibroblast growth factor 21. American Journal of Physiology-Endocrinology and Metabolism. 2021 Aug 03; 321:2, E292-E304.
- Qiu H, Song E, Hu Y, et al. Hepatocyte-Secreted Autotaxin Exacerbates Nonalcoholic Fatty Liver Disease Through Autocrine Inhibition of the PPARα/FGF21 Axis. Cellular and Molecular Gastroenterology and Hepatology. 2022 Aug:S2352-345X(22)00170-9
- Yu J, Laybuttab DR, Youngson NA, Morris MJ. Concurrent betaine administration enhances exercise-induced improvements to glucose handling in obese mice. Nutrition, Metabolism and Cardiovascular Diseases. 2022 Aug 18.