| Field | Specification |
|---|---|
| Target | |
| Alternative names | Fibroblast growth factor 21;FGF-21;FGF21;UNQ3115/PRO10196; |
| Gene ID | |
| UniProt # | |
| Reactivity | |
| Applications | |
| Immunogen | Expression system for standard: E.coli; Immunogen sequence: H29-S209 |
| Expression system | |
| Expression region | |
| Assay type | |
| Sample type(s) | Cell Culture Supernatant, Cell Lysate, Serum, Plasma |
| Sensitivity | |
| Detection range | |
| Assay time | |
| Storage | |
| Catalog no. (Mfr.) | |
| Main SKU |
Assay Principle
This sandwich ELISA quantifies human fibroblast growth factor 21 (gene FGF21) in cell culture supernatants, cell lysates, serum and plasma (heparin, EDTA). Strips are pre-coated with a capture antibody. After the sample is added, a biotinylated detection antibody binds the captured protein, and an avidin–biotin–peroxidase complex supplies the enzyme. TMB develops color in proportion to the bound target; the reaction is stopped and read at 450 nm, and concentrations are read from the standard curve.
Fibroblast growth factor 21 (FGF21) is an endocrine FGF made mainly by the liver in response to fasting and nutritional stress. It signals through FGF receptors together with the coreceptor β-Klotho to increase glucose uptake in fat cells and improve insulin sensitivity. FGF21 is widely studied in energy metabolism, obesity and metabolic disease.
Kit Components
| Anti-Human FGF21 Pre-coated 96-well strip microplate | 1 plate (96 wells) |
|---|---|
| Human FGF21 Standard | 2 vials, 10 ng/tube |
| Human FGF21 Biotinylated antibody (100x) | 100 µL |
| Avidin-Biotin-Peroxidase Complex (100x) | 100 µL |
| Sample Diluent | 30 mL |
| Antibody Diluent | 12 mL |
| Avidin-Biotin-Peroxidase Diluent | 12 mL |
| Color Developing Reagent (TMB) | 10 mL |
| Stop Solution | 10 mL |
| Wash Buffer (25x) | 20 mL |
| Adhesive plate sealers | 4 |
Also required (not supplied): microplate reader for 450 nm; incubator; automated plate washer (optional); precision pipettes for 0.5 µL through 1 mL volumes of aqueous solutions; multichannel pipettes (recommended for many samples); deionized or distilled water; 500 mL graduated cylinders; test tubes for dilution.
Kit components are not sold separately.
Performance Data
| Species | Human |
|---|---|
| Sample types | Cell culture supernatants, cell lysates, serum and plasma (heparin, EDTA) |
| Assay range | 31.2 pg/mL–2,000 pg/mL |
| Sensitivity | <10 pg/mL |
| Standard | Recombinant human FGF21 (His29–Ser209), expressed in E. coli |
| Cross-reactivity | None detected against related proteins in supplier testing |
| Intra-assay CV | 6.0–6.6% (3 samples, n = 16 each) |
| Inter-assay CV | 6.3–8.5% (3 samples, n = 24 each) |
| Format | 96 wells, removable strips |
Sample dilution: start at 1:1. In the supplier’s tests, serum and plasma samples measured about 20 pg/mL.
Example standard curve (OD450, pg/mL): 0 = 0.004; 31.2 = 0.065; 62.5 = 0.126; 125 = 0.222; 250 = 0.415; 500 = 0.822; 1000 = 1.548; 2000 = 2.579. Run a new standard curve on every plate.
Storage & Handling
Store at 4 °C for up to 6 months or −20 °C for up to 12 months. Avoid repeated freeze–thaw cycles. Ships on gel ice and tolerates up to 3 days at room temperature; freeze on arrival.
Safety
Sample data
| Concentration (pg/ml) | 0 | 31.2 | 62.5 | 125 | 250 | 500 | 1000 | 2000 |
| O.D. | 0.004 | 0.065 | 0.126 | 0.222 | 0.415 | 0.822 | 1.548 | 2.579 |
Intra/inter assay consistency
| Intra-Assay Precision | Inter-Assay Precision | |||||
|---|---|---|---|---|---|---|
| Sample | 1 | 2 | 3 | 1 | 2 | 3 |
| n | 16 | 16 | 16 | 24 | 24 | 24 |
| Mean (pg/ml) | 53 | 203 | 863 | 48 | 217 | 789 |
| Standard deviation | 3.23 | 12.18 | 56.95 | 3.4 | 13.67 | 67.06 |
| CV (%) | 6.1% | 6% | 6.6% | 7.1% | 6.3% | 8.5% |
Kit components
Description|Quantity Pre-coated 96-well strip microplate|1 Standard|2 vials Biotinylated antibody (100x)|100ul Avidin-Biotin-Peroxidase Complex (100x)|100ul Sample Diluent|30ml Antibody Diluent|12ml Avidin-Biotin-Peroxidase Diluent|12ml Color Developing Reagent (TMB)|10ml Stop Solution|10ml Wash Buffer (25x)|20ml Adhesive plate sealers|4Materials required but not provided
- Microplate Reader capable of reading absorbance at 450nm.
- Incubator.
- Automated plate washer (optional).
- Pipettes and pipette tips capable of precisely dispensing 0.5 µl through 1 ml volumes of aqueous solutions.
- Multichannel pipettes are recommended for large amount of samples.
- Deionized or distilled water.
- 500ml graduated cylinders.
- Test tubes for dilution.
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Ming-Fen Ho et al. (2022) Genome-wide association study for circulating FGF21 in patients with alcohol use disorder: Molecular links between the SNHG16 locus and catecholamine metabolism. Molecular Metabolism. 10.1016/j.molmet.2022.101534
Ebrahim Banitalebi et al. (2018) Effects of sprint interval or combined aerobic and resistance training on myokines in overweight women with type 2 diabetes: A randomized controlled trial. Life Sciences. 10.1016/j.lfs.2018.11.062
Kahraman S. et al. (2018) Association of serum betatrophin with fibroblast growth factor-21 in women with polycystic ovary syndrome. Journal of Endocrinological Investigation. 10.1007/s40618-018-0831-2
Aslıhan Ağaçdiken et al. (2023) Fluctuations of serum fibroblast growth factor-21 levels in prolonged fasting in male adults. Clinical Nutrition ESPEN. 10.1016/j.clnesp.2023.06.024
Liu Xinlei et al. (2022) Alternate-day fasting alleviates high fat diet induced non-alcoholic fatty liver disease through controlling PPARα/Fgf21 signaling. Molecular Biology Reports. 10.1007/s11033-022-07142-5
Erdem Nur Bengü et al. (2024) Effects of low fat diet on inflammatory parameters in individuals with obesity/overweight and non-alcoholic fatty liver disease: A cross-sectional study. Medicine. 10.1097/MD.0000000000037716
