Human bFGF/FGF2 (Basic Fibroblast Growth Factor) CLIA Kit (HUES00347)
- SKU:
- HUES00347
- Product Type:
- ELISA Kit
- ELISA Type:
- CLIA Kit
- Size:
- 96 Assays
- Sensitivity:
- 7.5pg/mL
- Range:
- 12.5-800pg/mL
- ELISA Type:
- Sandwich
- Synonyms:
- FGF-2, B-FGF, BFGF, FGFB, HBGF-2, prostatropin, heparin-binding growth factor 2
- Reactivity:
- Human
- Sample Type:
- Serum, plasma and other biological fluids
- Research Area:
- Cardiovascular
Description
| Assay type: | Sandwich |
| Format: | 96T |
| Assay time: | 4.5h |
| Reactivity: | Human |
| Detection method: | Chemiluminescence |
| Detection range: | 12.50-800 pg/mL |
| Sensitivity: | 7.50 pg/mL |
| Sample volume: | 100µL |
| Sample type: | Serum, plasma and other biological fluids |
| Repeatability: | CV < 15% |
| Specificity: | This kit recognizes Human bFGF/FGF2 in samples. No significant cross-reactivity or interference between Human bFGF/FGF2 and analogues was observed. |
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 bFGF/FGF2. 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 bFGF/FGF2 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 bFGF/FGF2, 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 bFGF/FGF2. The concentration of Human bFGF/FGF2 in the samples can be calculated by comparing the RLU of the samples to the standard curve.
| UniProt Protein Function: | FGF2: Plays an important role in the regulation of cell survival, cell division, angiogenesis, cell differentiation and cell migration. Functions as potent mitogen in vitro. Monomer. Homodimer. Interacts with FGFR1, FGFR2, FGFR3 and FGFR4. Affinity between fibroblast growth factors (FGFs) and their receptors is increased by heparan sulfate glycosaminoglycans that function as coreceptors. Interacts with CSPG4, FGFBP1 and TEC. Found in a complex with FGFBP1, FGF1 and FGF2. Expressed in granulosa and cumulus cells. Expressed in hepatocellular carcinoma cells, but not in non- cancerous liver tissue. Belongs to the heparin-binding growth factors family. 4 isoforms of the human protein are produced by alternative initiation. |
| UniProt Protein Details: | Protein type:Activator; Motility/polarity/chemotaxis Chromosomal Location of Human Ortholog: 4q26 Cellular Component: extracellular space; extracellular region; nucleus Molecular Function:heparin binding; protein binding; ligand-dependent nuclear receptor transcription coactivator activity; growth factor activity; cytokine activity; chemoattractant activity; fibroblast growth factor receptor binding Biological Process: extracellular matrix organization and biogenesis; wound healing; nerve growth factor receptor signaling pathway; somatic stem cell maintenance; activation of MAPK activity; positive regulation of transcription, DNA-dependent; chemotaxis; signal transduction; hyaluronan catabolic process; growth factor dependent regulation of satellite cell proliferation; positive regulation of MAP kinase activity; positive chemotaxis; positive regulation of cell proliferation; embryonic morphogenesis; positive regulation of cardiac muscle cell proliferation; epidermal growth factor receptor signaling pathway; nervous system development; phosphoinositide-mediated signaling; fibroblast growth factor receptor signaling pathway; positive regulation of cell fate specification; positive regulation of blood vessel endothelial cell migration; positive regulation of phosphoinositide 3-kinase activity; negative regulation of blood vessel endothelial cell migration; regulation of angiogenesis; positive regulation of angiogenesis; organ morphogenesis; cell migration during sprouting angiogenesis; ureteric bud branching; release of sequestered calcium ion into cytosol; positive regulation of cell division; Ras protein signal transduction; insulin receptor signaling pathway; phosphatidylinositol biosynthetic process; innate immune response; positive regulation of transcription from RNA polymerase II promoter; positive regulation of endothelial cell proliferation; inositol phosphate biosynthetic process |
| NCBI Summary: | The protein encoded by this gene is a member of the fibroblast growth factor (FGF) family. FGF family members bind heparin and possess broad mitogenic and angiogenic activities. This protein has been implicated in diverse biological processes, such as limb and nervous system development, wound healing, and tumor growth. The mRNA for this gene contains multiple polyadenylation sites, and is alternatively translated from non-AUG (CUG) and AUG initiation codons, resulting in five different isoforms with distinct properties. The CUG-initiated isoforms are localized in the nucleus and are responsible for the intracrine effect, whereas, the AUG-initiated form is mostly cytosolic and is responsible for the paracrine and autocrine effects of this FGF. [provided by RefSeq, Jul 2008] |
| UniProt Code: | P09038 |
| NCBI GenInfo Identifier: | 261260095 |
| NCBI Gene ID: | 2247 |
| NCBI Accession: | P09038. 3 |
| UniProt Secondary Accession: | P09038,O00527, P78443, Q16443, Q5PY50, Q7KZ11, Q7KZ72 Q9UC54, Q9UCS5, Q9UCS6, A4LBB8, |
| UniProt Related Accession: | P09038 |
| Molecular Weight: | |
| NCBI Full Name: | Fibroblast growth factor 2 |
| NCBI Synonym Full Names: | fibroblast growth factor 2 (basic) |
| NCBI Official Symbol: | FGF2 |
| NCBI Official Synonym Symbols: | BFGF; FGFB; FGF-2; HBGF-2 |
| NCBI Protein Information: | fibroblast growth factor 2; prostatropin; heparin-binding growth factor 2; basic fibroblast growth factor bFGF |
| UniProt Protein Name: | Fibroblast growth factor 2 |
| UniProt Synonym Protein Names: | Basic fibroblast growth factor; bFGF; Heparin-binding growth factor 2; HBGF-2 |
| Protein Family: | Fibroblast growth factor |
| UniProt Gene Name: | FGF2 |
| UniProt Entry Name: | FGF2_HUMAN |
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 |
| 800 | 50746 54538 | 52642 | 52616 |
| 400 | 21274 22194 | 21734 | 21708 |
| 200 | 9867 9549 | 9708 | 9682 |
| 100 | 4434 4670 | 4552 | 4526 |
| 50 | 2323 2053 | 2188 | 2162 |
| 25 | 1063 1057 | 1060 | 1034 |
| 12.50 | 489 529 | 509 | 483 |
| 0 | 25 27 | 26 | -- |
Precision
Intra-assay Precision (Precision within an assay): 3 samples with low, mid range and high level Human bFGF/FGF2 were tested 20 times on one plate, respectively.
Inter-assay Precision (Precision between assays): 3 samples with low, mid range and high level Human bFGF/FGF2 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) | 39.16 | 84.03 | 280.73 | 39.06 | 77.90 | 290.55 |
| Standard deviation | 4.97 | 5.98 | 32.06 | 4.66 | 8.37 | 20.34 |
| C V (%) | 12.69 | 7.12 | 11.42 | 11.93 | 10.74 | 7.00 |
Recovery
The recovery of Human bFGF/FGF2 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) | 91-103 | 97 |
| EDTA plasma (n=5) | 98-115 | 105 |
| Cell culture media (n=5) | 99-117 | 107 |
Linearity
Samples were spiked with high concentrations of Human bFGF/FGF2 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) | ||
| 1:2 | Range (%) | 93-107 | 85-100 | 90-106 |
| Average (%) | 99 | 91 | 97 | |
| 1:4 | Range (%) | 97-112 | 94-109 | 93-106 |
| Average (%) | 105 | 101 | 98 | |
| 1:8 | Range (%) | 94-108 | 99-113 | 93-107 |
| Average (%) | 100 | 105 | 99 | |
| 1:16 | Range (%) | 87-102 | 96-113 | 96-111 |
| Average (%) | 95 | 104 | 102 |
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 ×12 strips | -20°C, 6 months |
| Reference Standard | 2 vials | |
| Concentrated Biotinylated Detection Ab (100×) | 1 vial, 120 µL | |
| Concentrated HRP Conjugate (100×) | 1 vial, 120 µL | -20°C(shading light), 6 months |
| Reference Standard & Sample Diluent | 1 vial, 20 mL | 4°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 A | 1 vial, 5 mL | 4°C (shading light) |
| Substrate Reagent B | 1 vial, 5 mL | 4°C (shading light) |
| Plate Sealer | 5 pieces | |
| Product Description | 1 copy | |
| Certificate of Analysis | 1 copy |
- 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.
- Aliquot 100µl of standard solutions into the standard wells.
- Add 100µl of Sample / Standard dilution buffer into the control (zero) well.
- Add 100µl of properly diluted sample (serum, plasma, tissue homogenates and otherbiological fluids. ) into test sample wells.
- Cover the plate with the sealer provided in the kit and incubate for 90 min at 37°C.
- 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.
- 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.
- Add 100µL of HRP Conjugate working solution to each well. Cover with a plate seal andincubate for 30 min at 37°C.
- Aspirate or decant the solution from each well. Repeat the wash process for five times asconducted in step 7.
- 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.
- Determine the RLU value of each well immediately.
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