The ImmunoDiangostics Human FGF-19 Immunoassay is a Solid Phase ELISA that is designed to accurately detect FGF-19 in human serum, plasma, and cell culture supernatant samples.
The ImmunoDiagnostics Fibroblast Growth Factor-19 (FGF-19) ELISA is a reliable and reproducible tool for the quantitative detection of FGF-19 antigen. The assay is intended for use on human serum, plasma, or cell culture supernatants.
Numerous application fields exist, but particularly:
- Metabolic Syndrome: Subjects with metabolicy syndrome, non-alcoholic fatty liver disease (NAFLD), and insulin resistance have depressed levels of FGF-19.
- Oncology: FGF-19 is found in roughly half of hepatocellular carcinoma cell lines and was associated with larger and more aggressive growth.
FGF-19 Antibody Coated Microstrips: One microplate, 12 strips with 8 wells each, 96 dry wells in total. The wells are coated with a polyclonal anti- human FGF-19 antibody. The microplate is sealed in a foil bag. Any unused strips should be returned to the bag and resealed for future use.
Detection Antibody Solution: Concentrate (100x conc). One vial containing 0.12 mL of a biotin conjugated mouse polyclonal anti- human FGF-19. Detection antibody should be diluted with only the 1X Assay Buffer needed.
Assay Buffer: Concentrate (5x conc). One vial containing 20 mL of buffer for dilution of the detection antibody. If precipitates are observed in the concentrated buffer, warm at 37oC until the precipitates disappear. Dilute 20 mL of 5x Assay Buffer with 80 mL diH2O to make a 1x concentration prior to use.
Human FGF-19 Standard: 2000 pg of recombinant human FGF-19 in a buffered protein base, lyophilized. Reconstitute with 1 mL 1x Assay Buffer to prepare 2000 pg/mL standard stock. Prepare a serial dilution series with 1x Assay Buffer. The reconstituted standard stock can be aliquoted and stored at -20oC for one month. Avoid repeated freeze/thaw cycles.
Wash Buffer: Concentrate (10x conc). One vial containing 50 mL of 10x wash buffer. If precipitates are observed in the 10x solution, warm at 37oC until the precipitates disappear. Prepare 1x Wash Buffer my mixing 50 mL of 10x Wash Buffer with 450 mL diH2O.
STP-HRP Solution: Concentrate (200x) solution. One vial containing 0.06 mL of 200x solution. Prepare only as much 1x solution as needed.
Substrate Solution: One bottle containing 12 mL of TMB (3,3’,5,5’-Tetramethylbenzidine) substrate. Ready-to-use. Do not expose to light.
Stop Solution: One vial containing 12 mL of ready-to-use stop solution.
The FGF-19 ELISA is a quantitative sandwich enzyme immunoassay. Standard, samples, and any desired controls are pipetted into the wells pre-coated with solid-phase capture antibody. After washing away any unbound substances, a biotin-labelled detection antibody, specific for human FGF-19, is added to the wells, and any unbound antibody is washed out. Streptavidin-HRP conjugate is then added and a TMB solution is added to develop color in proportion to the amount of human FGF-19 initially bound. The color development is stopped by addition of an acidifying solution and the optical density is determined. By plotting a standard curve versus measured absorbance, the amount of antigen in the sample can be calculated. The concentration of the antigen is expressed in pg/mL.
The kit contains calibrators and can be used in any laboratory equipped with a plate reader. The kit is optimized for the quantitative determination of human FGF-19 concentrations in serum plasma, and cell culture supernatant samples. The test can provide results within 3 hours.
Example only! Do not use for calculations
4 Parameter Curve
Fibroblast Growth Factor-19 (FGF-19) is a member of the subfamily of FGFs that include both FGF-21 and FGF-23. Each member of this subfamily has important functions in the regulation of nutrient metabolism. FGF-19 is synthesized in the ileum in response to bile acids in the intestines and acts as a hormone to maintain proper bile acid homeostasis. Pharmacologic research has shown that FGF-19 can improve metabolic rate and lower serum glucose levels as well as hepatic triglycerides and cholesterol levels in hyperglycemic and obese animal models. Like insulin, FGF-19 functions as a postprandial hormone to govern hepatic protein synthesis, glycogen synthesis and gluconeogenesis, but not lipogenesis.
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