Human 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 pre-coated with a rabbit polyclonal antibody specific for human FGF-21. Standards and samples are pipetted into the wells and any human FGF-21 present is bound by the immobilized antibody. After washing away any unbound substances, a biotin labelled polyclonal antibody specific for human 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 human 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 human 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.
Human FGF-21 (pg/mL)
The lowest level of human FGF-21 that can be detected by this assay is 30 pg/mL.
Cross Reactivity of recombinant proteins
Analyte Cross Reactivity Mouse FGF-21
Human FABP4 No Human LCN2 No Human Adiponectin No Human ANGPL4 No
Intra-assay Precision (Precision within an assay)
Two samples of known concentration were tested 12 times on one plate.
Sample Mean (pg/mL) SD (pg/mL) CV (%) 1 93.3 3.72 4.0 2 547 27.6 5.0
Inter-assay Precision (Precision between assays)
Two samples of known concentration were tested in 10 separate assays.
Sample Mean (pg/mL) SD (pg/mL) CV (%) 1 273.5 27.9 10.2 2 335 11.8 3.5
To assess the linearity of the assay, samples containing and/or spiked with high concentrations of human FGF-21 were serially diluted with the 1×Assay buffer to produce samples with values within the dynamic range of the assay.
Dilution Mean (pg/mL) Expected (pg/mL) Recovery (%) Neat (non-diluted) 448 448 100 1:2 245 224 109 1:4 123 112 109 1:8 63.6 56 113
Dilution Mean (pg/mL) Expected (pg/mL) Recovery (%) Neat (non-diluted) 318 318 100 1:2 170 159 106 1:4 84.3 79.5 106 1:8 43.1 39.75 108
PUBLICATIONS CITING THIS PRODUCT
- Ong KL, Rye KA, O'Connell R, Jenkins AJ, Brown C, Xu A, Sullivan DR, Barter PJ, Keech AC, FIELD Study Investigators. Long-term fenofibrate therapy increases fibroblast growth factor 21 and retinol-binding protein 4 in subjects with type 2 diabetes. The Journal of Clinical Endocrinology & Metabolism. 2012 Dec 1;97(12):4701-8.
- Li H, Dong K, Fang Q, Hou X, Zhou M, Bao Y, Xiang K, Xu A, Jia W. High serum level of fibroblast growth factor 21 is an independent predictor of non-alcoholic fatty liver disease: a 3-year prospective study in China. Journal of hepatology. 2013 Mar 1;58(3):557-63.
- Heilbronn LK, Campbell LV, Xu A, Samocha-Bonet D. Metabolically protective cytokines adiponectin and fibroblast growth factor-21 are increased by acute overfeeding in healthy humans. PloS one. 2013 Oct 18;8(10):e78864.
- Shen Y, Ma X, Zhou J, Pan X, Hao Y, Zhou M, Lu Z, Gao M, Bao Y, Jia W. Additive relationship between serum fibroblast growth factor 21 level and coronary artery disease. Cardiovascular diabetology. 2013 Dec;12(1):1-7.
- Ong KL, Januszewski AS, O’Connell R, Jenkins AJ, Xu A, Sullivan DR, Barter PJ, Hung WT, Scott RS, Taskinen MR, Keech AC. The relationship of fibroblast growth factor 21 with cardiovascular outcome events in the Fenofibrate Intervention and Event Lowering in Diabetes study. Diabetologia. 2015 Mar;58(3):464-73.
- Zhang W, Chu S, Ding W, Wang F. Serum level of fibroblast growth factor 21 is independently associated with acute myocardial infarction. PLoS One. 2015 Jun 19;10(6):e0129791.
- Besseling J, Ong KL, Huijgen R, Rye KA, Hovingh GK, Hutten BA, Lambert G. The impact of LDLR function on fibroblast growth factor 21 levels. Atherosclerosis. 2015 Aug 1;241(2):322-5.
- Ong KL, Januszewski AS, O’Connell R, Buizen L, Jenkins AJ, Xu A, Sullivan DR, Barter PJ, Scott RS, Taskinen MR, Rye KA. Relationship of fibroblast growth factor 21 with baseline and new on-study microvascular disease in the Fenofibrate Intervention and Event Lowering in Diabetes study. Diabetologia. 2015 Sep;58(9):2035-44.
- Chen DL, Liess C, Poljak A, Xu A, Zhang J, Thoma C, Trenell M, Milner B, Jenkins AB, Chisholm DJ, Samocha-Bonet D. Phenotypic characterization of insulin-resistant and insulin-sensitive obesity. The Journal of Clinical Endocrinology & Metabolism. 2015 Nov 1;100(11):4082-91.
- Xiao Y, Liu L, Xu A, Zhou P, Long Z, Tu Y, Chen X, Tang W, Huang G, Zhou Z. Serum fibroblast growth factor 21 levels are related to subclinical atherosclerosis in patients with type 2 diabetes. Cardiovascular diabetology. 2015 Dec;14(1):1-8.
- Ye D, Li H, Wang Y, Jia W, Zhou J, Fan J, Man K, Lo C, Wong C, Wang Y, Lam KS. Circulating fibroblast growth factor 21 is a sensitive biomarker for severe ischemia/reperfusion injury in patients with liver transplantation. Scientific reports. 2016 Jan 25;6(1):1-9.
- Woo YC, Lee CH, Fong CH, Xu A, Tso AW, Cheung BM, Lam KS. Serum fibroblast growth factor 21 is a superior biomarker to other adipokines in predicting incident diabetes. Clinical endocrinology. 2017 Jan;86(1):37-43.
- Chen DL, Brown R, Liess C, Poljak A, Xu A, Zhang J, Trenell M, Jenkins A, Chisholm D, Samocha-Bonet D, Macefield VG. Muscle sympathetic nerve activity is associated with liver insulin sensitivity in obese non-diabetic men. Frontiers in physiology. 2017 Feb 28;8:101.
- Lee CH, Woo YC, Chow WS, Cheung CY, Fong CH, Yuen MM, Xu A, Tse HF, Lam KS. Role of circulating fibroblast growth factor 21 measurement in primary prevention of coronary heart disease among Chinese patients with type 2 diabetes mellitus. Journal of the American Heart Association. 2017 Jun 6;6(6):e005344.
- Yang K, Deng HB, Man AW, Song E, Zhang J, Luo C, Cheung BM, Yuen KY, Jensen PS, Irmukhamedov A, Elie AG. Measuring non‐polyaminated lipocalin‐2 for cardiometabolic risk assessment. ESC heart failure. 2017 Nov;4(4):563-75.
- Shen Y, Zhang X, Pan X, Xu Y, Xiong Q, Lu Z, Ma X, Bao Y, Jia W. Contribution of serum FGF21 level to the identification of left ventricular systolic dysfunction and cardiac death. Cardiovascular diabetology. 2017 Dec;16(1):1-7.
- Shen Y, Zhang X, Xu Y, Xiong Q, Lu Z, Ma X, Bao Y, Jia W. Serum FGF21 is associated with future cardiovascular events in patients with coronary artery disease. Cardiology. 2018;139(4):212-8.
- 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.
- Hui TH, McClelland RL, Allison MA, Rodriguez CJ, Kronmal RA, Heckbert SR, Michos ED, Barter PJ, Rye KA, Ong KL. The relationship of circulating fibroblast growth factor 21 levels with incident atrial fibrillation: The Multi-Ethnic Study of Atherosclerosis. Atherosclerosis. 2018 Feb 1;269:86-91.
- Liu JJ, Liu S, Choo RW, Wee SL, Xu A, Lim SC. Sex modulates the association of fibroblast growth factor 21 with end‐stage renal disease in Asian people with Type 2 diabetes: a 6.3‐year prospective cohort study. Diabetic Medicine. 2018 Jul;35(7):880-6.
- Wong YK, Cheung CY, Tang CS, Au KW, Hai JS, Lee CH, Lau KK, Cheung BM, Sham PC, Xu A, Lam KS. Age-biomarkers-clinical risk factors for prediction of cardiovascular events in patients with coronary artery disease. Arteriosclerosis, thrombosis, and vascular biology. 2018 Oct;38(10):2519-27.
- Hu Q, Wang C, Liu F, He J, Wang F, Wang W, You P. High serum levels of FGF21 are decreased in bipolar mania patients during psychotropic medication treatment and are associated with increased metabolism disturbance. Psychiatry research. 2019 Feb 1;272:643-8.
- Ong KL, Hui N, Januszewski AS, Kaakoush NO, Xu A, Fayyad R, DeMicco DA, Jenkins AJ, Keech AC, Waters DD, Barter PJ. High plasma FGF21 levels predicts major cardiovascular events in patients treated with atorvastatin (from the Treating to New Targets [TNT] Study). Metabolism. 2019 Apr 1;93:93-9.
- Tang A, Coster AC, Tonks KT, Heilbronn LK, Pocock N, Purtell L, Govendir M, Blythe J, Zhang J, Xu A, Chisholm DJ. Longitudinal changes in insulin resistance in normal weight, overweight and obese individuals. Journal of clinical medicine. 2019 May;8(5):623.
- Anuwatmatee S, Allison MA, Shlipak MG, McClelland RL, Kramer H, Tang S, Hou L, Rye KA, Ong KL. Relationship of fibroblast growth factor 21 with kidney function and albuminuria: multi-ethnic study of atherosclerosis. Nephrology Dialysis Transplantation. 2019 Jun 1;34(6):1009-16.
- Mak LY, Lee CH, Cheung KS, Wong DK, Liu F, Hui RW, Fung J, Xu A, Lam KS, Yuen MF, Seto WK. Association of adipokines with hepatic steatosis and fibrosis in chronic hepatitis B patients on long‐term nucleoside analogue. Liver International. 2019 Jul;39(7):1217-25.
- Harari A, Coster AC, Jenkins A, Xu A, Greenfield JR, Harats D, Shaish A, Samocha-Bonet D. Obesity and insulin resistance are inversely associated with serum and adipose tissue carotenoid concentrations in adults. The Journal of nutrition. 2020 Jan 1;150(1):38-46.
- Liu Y, Wang Y, Ni Y, Cheung CK, Lam KS, Wang Y, Xia Z, Ye D, Guo J, Tse MA, Panagiotou G. Gut microbiome fermentation determines the efficacy of exercise for diabetes prevention. Cell metabolism. 2020 Jan 7;31(1):77-91.
- Tucker B, McClelland RL, Allison MA, Budoff MJ, Wu BJ, Barter PJ, Rye KA, Ong KL. Relationship of fibroblast growth factor 21 levels with inflammation, lipoproteins and non-alcoholic fatty liver disease. Atherosclerosis. 2020 Apr 1;299:38-44.
- Lui DT, Lee CH, Chau VW, Fong CH, Yeung KM, Lam JK, Lee AC, Chow WS, Tan KC, Woo YC, Lam KS. Potential role of fibroblast growth factor 21 in the deterioration of bone quality in impaired glucose tolerance. Journal of Endocrinological Investigation. 2020 Jun 29:1-8.
- Wu L, Qian L, Zhang L, Zhang J, Zhou J, Li Y, Hou X, Fang Q, Li H, Jia W. Fibroblast Growth Factor 21 is Related to Atherosclerosis Independent of Nonalcoholic Fatty Liver Disease and Predicts Atherosclerotic Cardiovascular Events. Journal of the American Heart Association. 2020 Jun 2;9(11):e015226.