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Human FGF21(Fibroblast Growth Factor 21) ELISA Kit

Citations(8)Uniprot : Q9NSA1

Cat.No.:E-EL-H0074
Reactivity: Human

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Product Details

Properties

Assay type	Sandwich-ELISA
Format	96T/48T
Assay time	3.5h
Detection range	31.25-2000 pg/mL
Sensitivity	18.75 pg/mL
Sample type &Sample volume	serum, plasma and other biological fluids; 100μL
Specificity	This kit recognizes Human FGF21 in samples. No significant cross-reactivity or interference between Human FGF21 and analogues was observed.
Reproducibility	Both intra-CV and inter-CV are < 10%.
Application	This ELISA kit applies to the in vitro quantitative determination of Human FGF21 concentrations in serum, plasma and other biological fluids.

Test Principle

This ELISA kit uses the Sandwich-ELISA principle. The micro ELISA plate provided in this kit has been pre-coated with an antibody specific to Human FGF21. Standards or samples are added to the micro ELISA plate wells and combined with the specific antibody. Then a biotinylated detection antibody specific for Human FGF21 and Avidin-Horseradish Peroxidase (HRP) conjugate are added successively to each micro plate well and incubated. Free components are washed away. The substrate solution is added to each well. Only those wells that contain Human FGF21, biotinylated detection antibody and Avidin-HRP conjugate will appear blue in color. The enzyme-substrate reaction is terminated by the addition of stop solution and the color turns yellow. The optical density (OD) is measured spectrophotometrically at a wavelength of 450 nm ± 2 nm. The OD value is proportional to the concentration of Human FGF21. You can calculate the concentration of Human FGF21 in the samples by comparing the OD of the samples to the standard curve.

Kit components & Storage

An unopened kit can be stored at 2-8℃ for 1 month. If the kit is not supposed to be used within 1 month, store the components separately according to the following conditions once the kit is received.

Item	Specifications	Storage
Micro ELISA Plate(Dismountable)	96T: 8 wells ×12 strips 48T: 8 wells ×6 strips	-20℃, 6 months
Reference Standard	96T: 2 vials 48T: 1 vial
Concentrated Biotinylated Detection Ab (100×)	96T: 1 vial, 120 μL 48T: 1 vial, 60 μL
Concentrated HRP Conjugate (100×)	96T: 1 vial, 120 μL 48T: 1 vial, 60 μL	-20℃(Protect from light), 6 months
Reference Standard & Sample Diluent	1 vial, 20 mL	2-8°C, 6 months
Biotinylated Detection Ab Diluent	1 vial, 14 mL
HRP Conjugate Diluent	1 vial, 14 mL
Concentrated Wash Buffer (25×)	1 vial, 30 mL
Substrate Reagent	1 vial, 10 mL	2-8℃(Protect from light)
Stop Solution	1 vial, 10 mL	2-8°C
Plate Sealer	5 pieces
Manual	1 copy
Certificate of Analysis	1 copy

Technical Data

Precision

Intra-assay Precision (Precision within an assay): 3 samples with low, mid range and high level Human FGF21 were tested 20 times on one plate, respectively.
Inter-assay Precision (Precision between assays): 3 samples with low, mid range and high level Human FGF21 were tested on 3 different plates, 20 replicates in each plate.

	Intra-assay Precision			Inter-assay Precision
Sample	1	2	3	1	2	3
n	20	20	20	20	20	20
Mean (pg/mL)	97.95	240.13	857.10	94.53	241.91	786.68
Standard deviation	6.34	9.99	42.17	6.33	10.21	33.20
CV (%)	6.47	4.16	4.92	6.70	4.22	4.22

Recovery

The recovery of Human FGF21 spiked at three different levels in samples throughout the range of the assay was evaluated in various matrices.

Sample Type	Range (%)	Average Recovery (%)
Serum(n=8)	84-95	90
EDTA plasma (n=8)	84-98	90
Cell culture media (n=8)	88-102	94

Linearity

Samples were spiked with high concentrations of Human FGF21 and diluted with Reference Standard & Sample Diluent to produce samples with values within the range of the assay.

Target Information

Database Links	SwissProt: Q9NSA1
Synonyms	FGF-21
Research Area	Cancer, Cardiovascular, Signal transduction

Assay Procedures

	1. Add 100μL standard or sample to the wells. Incubate for 90 min at 37°C
	2. Discard the liquid, immediately add 100μL Biotinylated Detection Ab working solution to each well. Incubate for 60 min at 37°C
	3. Aspirate and wash the plate for 3 times
	4. Add 100μL HRP conjugate working solution. Incubate for 30 min at 37°C. Aspirate and wash the plate for 5 times
	5. Add 90μL Substrate Reagent. Incubate for 15 min at 37°C
	6. Add 50μL Stop Solution
	7. Read the plate at 450nm immediately. Calculation of the results

Citations

Antioxidants (2020) IF: 5.014
Oxidative Stress and Pro-Inflammatory Status in Patients with Non-Alcoholic Fatty Liver Disease

DOI: 10.3390/antiox9080759

Sample: plasma
Frontiers in Pharmacology (2021) IF: 5.811
Fibroblast Growth Factor 21 Ameliorates Na_V1.5 and Kir2.1 Channel Dysregulation in Human AC16 Cardiomyocytes.

DOI: 10.3389/fphar.2021.715466

Sample: serum
INTERNATIONAL JOURNAL OF MOLECULAR SCIENCES (2022) IF: 5.924
Pyruvate Upregulates Hepatic FGF21 Expression by Activating PDE and Inhibiting cAMP–Epac–CREB Signaling Pathway

DOI: 10.3390/ijms23105490

Sample: HepG2 cell
BASIC & CLINICAL PHARMACOLOGY & TOXICOLOGY (2022) IF: 3.688
α-Lipoic acid up-regulates gene expression but reduces protein levels of fibroblast growth factor 21 in HepG2 cells

DOI: 10.1111/bcpt.13775

PMID: 35838000
Sample: HepG2 cell
Diagnostics (2021) IF: 3.706
Diagnostic Performance of Serum Biomarkers Fibroblast Growth Factor 21, Galectin-3 and Copeptin for Heart Failure with Preserved Ejection Fraction in a Sample of Patients with Type 2 Diabetes Mellitus

DOI: 10.3390/diagnostics11091577

PMID: 34573919
Sample: serum
Geriatrics & Gerontology International (2021) IF: 2.73
Association of FGF-19 and FGF-21 levels with primary sarcopenia

DOI: 10.1111/ggi.14263

PMID: 34405516
Sample: serum
JOURNAL OF DIABETES AND ITS COMPLICATIONS (2021) IF: 2.852
Effects of increased physical activity and/or weight loss diet on serum myokine and adipokine levels in overweight adults with impaired glucose metabolism

DOI: 10.1016/j.jdiacomp.2021.107892

PMID: 33685795
Sample: Serum
Open Life Sciences (2022) IF: 1.311
β-Hydroxybutyrate upregulates FGF21 expression through inhibition of histone deacetylases in hepatocytes

DOI: 10.1515/biol-2022-0095

Sample: HepG2 cell

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Reviews/Q&A

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Reviews
Q&A

I******************gSubmitted [ Nov 28 2023 ]

Asked: Hi, I am a medical doctor, a part 2 resident of the department of Chemical Pathology UCTH, Calabar, Nigeria . I intend to use this product for my research project and my supervisor wants me to furnish him with information and with references on how I am going to process and preserve my samples. He is particularly interested on the stability of fibroblast growth factor 21 in plasma or serum if stored at -80\'C. I\'ll be glad if this information can be made available to me. Moreover , the cost of the kit and delivery to me should be stated. Thanks as I look forward to hearing from you. Dr. Asanga.

adminSubmitted [ Nov 28 2023 ]

Answered: Dr. Asanga, Article No. E-EL-H0074, human Fibroblast Growth Factor 21. Serum or plasma should be centrifuged in time after extraction of whole blood sample, which can be stably stored at -80℃ for 3 months to avoid repeated freeze-thaw. We also sent an email to you ,please check .

D*********oSubmitted [ Feb 18 2021 ]

Asked: what what is the substrate and the stop solution used in this reagent?

adminSubmitted [ Feb 18 2021 ]

Answered: Substrate: TMB Stop solution: 1M sulfuric acid

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