Human FGF19 (Fibroblast Growth Factor 19) ELISA Kit (HUES01360)

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HUES01360
€539

Description

ELISA Kit Technical ManualMSDS

Human FGF19 (Fibroblast Growth Factor 19) ELISA Kit

This ELISA kit uses Sandwich-ELISA as the method. The micro ELISA plate provided in this kit has been pre-coated with an antibody specific to Human FGF19 . Standards or samples are added to the appropriate micro ELISA plate wells and combined with the specific antibody. Then a biotinylated detection antibody specific for Human FGF19 and Avidin-Horseradish Peroxidase (HRP) conjugate are added to each micro plate well successively and incubated. Free components are washed away. The substrate solution is added to each well. Only those wells that contain Human FGF19, biotinylated detection antibody and Avidin-HRP conjugate will appear blue in color. The enzyme-substrate reaction is terminated by adding 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 FGF19. The concentration of Human FGF19 in samples can be calculated by comparing the OD of the samples to the standard curve.

Assay typeSandwich
Format96T
Assay time4.5h
ReactivityHuman
Detection MethodColormetric
Detection Range15.63-1000 pg/mL
Sensitivity9.38 pg/mL
Sample Volume Required Per Well100uL
Sample TypeSerum, plasma and other biological fluids

Specificity

This kit recognizes Human FGF19 in samples. No significant cross-reactivity or interference between Human FGF19 and analogues was observed.

Typical Data

As the OD values of the standard curve may vary according to the conditions of the actual assay performance (e.g. operator, pipetting technique, washing technique or temperature effects), the operator should establish a standard curve for each test. Typical standard curve and data is provided below for reference only.

Concentration
(pg/mL)
O.D Average Corrected
10002.38
2.432
2.4062.339
5001.692
1.704
1.6981.631
2500.964
0.934
0.9490.882
1250.426
0.43
0.4280.361
62.50.253
0.239
0.2460.179
31.250.173
0.163
0.1680.101
15.630.113
0.125
0.1190.052
00.063
0.071
0.067--

Precision

Intra-assay Precision (Precision within an assay): 3 samples with low, mid range and high level Human FGF19 were tested 20 times on one plate, respectively.

Inter-assay Precision (Precision between assays): 3 samples with low, mid range and high level Human FGF19 were tested on 3 different plates, 20 replicates in each plate.

Intra-assay Precision Inter-assay Precision
Sample123123
n202020202020
Mean (pg/mL)48.71118.24371.8445.18125.53341.83
Standard deviation2.525.2714.762.816.6811.08
C V (%)5.174.463.976.225.323.24

Recovery

The recovery of Human FGF19 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=5)88-10295
EDTA plasma (n=5)93-109100
Cell culture media (n=5)84-9691

Linearity

Samples were spiked with high concentrations of Human FGF19 and diluted with Reference Standard & Sample Diluent to produce samples with values within the range of the assay.

Serum (n=5) EDTA plasma (n=5) Cell culture media (n=5)
1:2Range (%)94-10887-10091-101
Average (%)1009596
1:4Range (%)86-9982-9287-100
Average (%)938794
1:8Range (%)88-9879-9183-98
Average (%)938690
1:16Range (%)86-9982-9384-98
Average (%)938890

Kit Components & Storage

An unopened kit can be stored at 4'C for 1 month. If the kit is not used within 1 month, store the items separately according to the following conditions once the kit is received.

Item Specifications Storage
Micro ELISA Plate(Dismountable)8 wells X12 strips-20'C, 6 months
Reference Standard2 vials
Concentrated Biotinylated Detection Ab (100X)1 vial, 120 uL
Concentrated HRP Conjugate (100X)1 vial, 120 uL-20'C(shading light), 6 months
Reference Standard & Sample Diluent1 vial, 20 mL4'C, 6 months
Biotinylated Detection Ab Diluent1 vial, 14 mL
HRP Conjugate Diluent1 vial, 14 mL
Concentrated Wash Buffer (25X)1 vial, 30 mL
Substrate Reagent1 vial, 10 mL4'C(shading light)
Stop Solution1 vial, 10 mL4'C
Plate Sealer5 pieces
Product Description1 copy
Certificate of Analysis1 copy

Human FGF19 (Fibroblast Growth Factor 19) ELISA Kit (HUES01360) Assay procedure

    1. Set standard, test sample and control (zero) wells on the pre-coated plate and record theirpositions. It is recommended to measure each standard and sample in duplicate. Note: addall solutions to the bottom of the plate wells while avoiding contact with the well walls. Ensuresolutions do not foam when adding to the wells.
    2. Aliquot 100µl of standard solutions into the standard wells.
    3. Add 100µl of Sample / Standard dilution buffer into the control (zero) well.
    4. Add 100µl of properly diluted sample (serum, plasma, tissue homogenates and otherbiological fluids) into test sample wells.
    5. Cover the plate with the sealer provided in the kit and incubate for 90 min at 37°C.
    6. Aspirate the liquid from each well, do not wash. Immediately add 100µL of BiotinylatedDetection Ab working solution to each well. Cover the plate with a plate seal and gently mix.Incubate for 1 hour at 37°C.
    7. Aspirate or decant the solution from the plate and add 350µL of wash buffer to each welland incubate for 1-2 minutes at room temperature. Aspirate the solution from each well andclap the plate on absorbent filter paper to dry. Repeat this process 3 times. Note: a microplatewasher can be used in this step and other wash steps.
    8. Add 100µL of HRP Conjugate working solution to each well. Cover with a plate seal andincubate for 30 min at 37°C.
    9. Aspirate or decant the solution from each well. Repeat the wash process for five times asconducted in step 7.
    10. Add 90µL of Substrate Reagent to each well. Cover with a new plate seal and incubate forapproximately 15 min at 37°C. Protect the plate from light. Note: the reaction time can beshortened or extended according to the actual color change, but not by more than 30min.
    11. Add 50 µL of Stop Solution to each well. Note: Adding the stop solution should be done inthe same order as the substrate solution.
    12. Determine the optical density (OD value) of each well immediately with a microplate readerset at 450 nm.

Human FGF19 (Fibroblast Growth Factor 19) ELISA Kit (HUES01360) Protein Information

UniProt Protein Function:FGF19: Involved in the suppression of bile acid biosynthesis through down-regulation of CYP7A1 expression, following positive regulation of the JNK and ERK1/2 cascades. Stimulates glucose uptake in adipocytes. Activity requires the presence of KLB and FGFR4. Belongs to the heparin-binding growth factors family.
UniProt Protein Details:

Protein type:Motility/polarity/chemotaxis; Cytokine; Secreted, signal peptide; Secreted; Cell development/differentiation

Chromosomal Location of Human Ortholog: 11q13.1

Cellular Component: extracellular region

Molecular Function:protein binding; growth factor activity; fibroblast growth factor receptor binding

Biological Process: epidermal growth factor receptor signaling pathway; nervous system development; positive regulation of glucose import; fibroblast growth factor receptor signaling pathway; phosphoinositide-mediated signaling; nerve growth factor receptor signaling pathway; heart development; positive regulation of cell proliferation; insulin receptor signaling pathway; innate immune response; positive regulation of JNK cascade; positive regulation of protein amino acid phosphorylation; neural crest cell migration

NCBI Summary:The protein encoded by this gene is a member of the fibroblast growth factor (FGF) family. FGF family members possess broad mitogenic and cell survival activities, and are involved in a variety of biological processes including embryonic development cell growth, morphogenesis, tissue repair, tumor growth and invasion. This growth factor is a high affinity, heparin dependent ligand for FGFR4. Expression of this gene was detected only in fetal but not adult brain tissue. Synergistic interaction of the chick homolog and Wnt-8c has been shown to be required for initiation of inner ear development. [provided by RefSeq, Jul 2008]
UniProt Code:O95750
NCBI GenInfo Identifier:11132196
NCBI Gene ID:9965
NCBI Accession:O95750.1
UniProt Related Accession:O95750
Molecular Weight:216
NCBI Full Name:Fibroblast growth factor 19
NCBI Synonym Full Names:fibroblast growth factor 19
NCBI Official Symbol:FGF19  
NCBI Protein Information:fibroblast growth factor 19; FGF-19
UniProt Protein Name:Fibroblast growth factor 19
Protein Family:Fibroblast growth factor
UniProt Gene Name:FGF19  
UniProt Entry Name:FGF19_HUMAN
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Additional Information

Product type:
ELISA
Reactivity:
Human
ELISA Type:
Sandwich
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