Human FGF9 (Fibroblast Growth Factor 9) CLIA Kit (HUES00656)

SKU:
HUES00656
Product type:
ELISA
Reactivity:
Human
ELISA Type:
CLIA
€599

Description

ELISA Kit Technical ManualMSDS

Human FGF9 (Fibroblast Growth Factor 9) CLIA Kit

This kit uses Sandwich-CLIA as the method. The micro CLIA plate provided in this kit has been pre-coated with an antibody specific to Human FGF9 . Standards or samples are added to the appropriate micro CLIA plate wells and combined with the specific antibody. Then a biotinylated detection antibody specific for Human FGF9 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 FGF9, biotinylated detection antibody and Avidin-HRP conjugate will appear fluorescence. The Relative light unit (RLU) value is measured spectrophotometrically by the Chemiluminescence immunoassay analyzer. The RLU value is positively associated with the concentration of Human FGF9. The concentration of Human FGF9 in the samples can be calculated by comparing the RLU of the samples to the standard curve.

Assay typeSandwich
Format96T
Assay time4.5h
ReactivityHuman
Detection methodChemiluminescence
Detection range15.63-1000 pg/mL
Sensitivity9.38 pg/mL
Sample volume100uL
Sample typeSerum, plasma and other biological fluids
RepeatabilityCV < 15%

Specificity

This kit recognizes Human FGF9 in samples. No significant cross-reactivity or interference between Human FGF9 and analogues was observed.

Typical Data

As the RLU 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)
RLU Average Corrected
100048416
58934
5367553650
50021764
23214
2248922464
25010584
9838
1021110186
1254802
5000
49014876
62.52571
2335
24532428
31.251300
1262
12811256
15.63688
728
708683
025
25
25--

Precision

Intra-assay Precision (Precision within an assay): 3 samples with low, mid range and high level Human FGF9 were tested 20 times on one plate, respectively.

Inter-assay Precision (Precision between assays): 3 samples with low, mid range and high level Human FGF9 were tested on 3 different plates, 20 replicates in each plate.

  Intra-assay Precision Inter-assay Precision
Sample123123
n202020202020
Mean (pg/mL)48.57135.24447.2852.74139.87490.99
Standard deviation6.2914.8231.584.7411.9441.69
C V (%)12.9510.967.068.998.548.49

Recovery

The recovery of Human FGF9 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)100-113106
EDTA plasma (n=5)98-110103
Cell culture media (n=5)90-10497

Linearity

Samples were spiked with high concentrations of Human FGF9 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 (%)104-11891-10590-107
Average (%)1109697
1:4Range (%)94-11299-11489-101
Average (%)10210594
1:8Range (%)103-11796-11290-105
Average (%)10910296
1:16Range (%)84-97102-119102-117
Average (%)91109108

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 CLIA 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 Reagent A1 vial, 5 mL4'C (shading light)
Substrate Reagent B1 vial, 5 mL4'C (shading light)
Plate Sealer5 pieces
Product Description1 copy
Certificate of Analysis1 copy

Human FGF9 (Fibroblast Growth Factor 9) CLIA Kit (HUES00656) 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 100µL of Substrate mixture solution to each well. Cover with a new plate seal andincubate for no more than 5 min at 37°C. Protect the plate from light.
    11. Determine the RLU value of each well immediately.

Human FGF9 (Fibroblast Growth Factor 9) CLIA Kit (HUES00656) Protein Information

UniProt Protein Function:FGF9: Plays an important role in the regulation of embryonic development, cell proliferation, cell differentiation and cell migration. May have a role in glial cell growth and differentiation during development, gliosis during repair and regeneration of brain tissue after damage, differentiation and survival of neuronal cells, and growth stimulation of glial tumors. Defects in FGF9 are the cause of multiple synostoses syndrome type 3 (SYNS3). Multiple synostoses syndrome is an autosomal dominant condition characterized by progressive joint fusions of the fingers, wrists, ankles and cervical spine, characteristic facies and progressive conductive deafness. Belongs to the heparin-binding growth factors family.
UniProt Protein Details:

Protein type:Cytokine

Chromosomal Location of Human Ortholog: 13q11-q12

Cellular Component: extracellular space; cytoplasm; extracellular region; basement membrane

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

Biological Process: negative regulation of Wnt receptor signaling pathway; nerve growth factor receptor signaling pathway; embryonic skeletal development; negative regulation of transcription from RNA polymerase II promoter; signal transduction; positive regulation of vascular endothelial growth factor receptor signaling pathway; positive regulation of activin receptor signaling pathway; substantia nigra development; cell-cell signaling; positive regulation of MAPKKK cascade; protein import into nucleus; male sex determination; positive regulation of mesenchymal cell proliferation; positive regulation of cell proliferation; positive regulation of smoothened signaling pathway; chondrocyte differentiation; angiogenesis; regulation of timing of cell differentiation; embryonic gut development; positive regulation of cardiac muscle cell proliferation; embryonic limb morphogenesis; epidermal growth factor receptor signaling pathway; inner ear morphogenesis; fibroblast growth factor receptor signaling pathway; phosphoinositide-mediated signaling; male gonad development; osteoblast differentiation; positive regulation of cell division; insulin receptor signaling pathway; innate immune response; positive regulation of epithelial cell proliferation

Disease: Multiple Synostoses Syndrome 3

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 protein was isolated as a secreted factor that exhibits a growth-stimulating effect on cultured glial cells. In nervous system, this protein is produced mainly by neurons and may be important for glial cell development. Expression of the mouse homolog of this gene was found to be dependent on Sonic hedgehog (Shh) signaling. Mice lacking the homolog gene displayed a male-to-female sex reversal phenotype, which suggested a role in testicular embryogenesis. [provided by RefSeq, Jul 2008]
UniProt Code:P31371
NCBI GenInfo Identifier:544290
NCBI Gene ID:2254
NCBI Accession:P31371.3
UniProt Secondary Accession:P31371,Q3SY32, A8K427,
UniProt Related Accession:P31371
Molecular Weight:23,441 Da
NCBI Full Name:Fibroblast growth factor 9
NCBI Synonym Full Names:fibroblast growth factor 9
NCBI Official Symbol:FGF9  
NCBI Official Synonym Symbols:GAF; FGF-9; SYNS3; HBFG-9; HBGF-9  
NCBI Protein Information:fibroblast growth factor 9; heparin-binding growth factor 9; fibroblast growth factor 9 (glia-activating factor)
UniProt Protein Name:Fibroblast growth factor 9
UniProt Synonym Protein Names:Glia-activating factor; GAF; Heparin-binding growth factor 9; HBGF-9
Protein Family:Fibroblast growth factor
UniProt Gene Name:FGF9  
UniProt Entry Name:FGF9_HUMAN
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