Human AFGF/FGF1 (Acidic Fibroblast Growth Factor 1) CLIA Kit (HUES00069)
- SKU:
- HUES00069
- 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-1, ECGF, ECGF-beta, A-FGF, FGFA, ECGFA, ECGFB, FGF-Alpha, HBGF1, HBGF-1, ECGFB, GLIO703, Heparin-binding growth factor 1
- 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 AFGF/FGF1 in samples. No significant cross-reactivity or interference between Human AFGF/FGF1 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 AFGF/FGF1. 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 AFGF/FGF1 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 AFGF/FGF1, 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 AFGF/FGF1. The concentration of Human AFGF/FGF1 in the samples can be calculated by comparing the RLU of the samples to the standard curve.
| UniProt Protein Function: | FGF1: 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. Found in a complex with FGFBP1, FGF1 and FGF2. Interacts with FGFBP1. Part of a Cu(2+)-dependent multiprotein aggregate containing FGF1, S100A13 and SYT1. Interacts with SYT1. Interacts with S100A13. Belongs to the heparin-binding growth factors family. 2 isoforms of the human protein are produced by alternative splicing. |
| UniProt Protein Details: | Protein type:Motility/polarity/chemotaxis; Cell development/differentiation; Cytokine Chromosomal Location of Human Ortholog: 5q31 Cellular Component: nucleoplasm; extracellular space; proteinaceous extracellular matrix; nucleolus; extracellular region; cell cortex; cytosol Molecular Function:heparin binding; protein binding; growth factor activity; Hsp70 protein binding; fibroblast growth factor receptor binding Biological Process: epidermal growth factor receptor signaling pathway; anatomical structure morphogenesis; phosphoinositide-mediated signaling; fibroblast growth factor receptor signaling pathway; nerve growth factor receptor signaling pathway; multicellular organismal development; positive regulation of cholesterol biosynthetic process; signal transduction; positive regulation of MAP kinase activity; positive regulation of angiogenesis; induction of an organ; positive regulation of cell division; positive regulation of cell proliferation; insulin receptor signaling pathway; innate immune response; positive regulation of transcription from RNA polymerase II promoter; angiogenesis; positive regulation of epithelial cell proliferation; positive regulation of cell migration; lung development |
| 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 functions as a modifier of endothelial cell migration and proliferation, as well as an angiogenic factor. It acts as a mitogen for a variety of mesoderm- and neuroectoderm-derived cells in vitro, thus is thought to be involved in organogenesis. Multiple alternatively spliced variants encoding different isoforms have been described. [provided by RefSeq, Jan 2009] |
| UniProt Code: | P05230 |
| NCBI GenInfo Identifier: | 122737 |
| NCBI Gene ID: | 2246 |
| NCBI Accession: | P05230. 1 |
| UniProt Secondary Accession: | P05230,P07502, Q16588, B2R5T0, D3DQF2, |
| UniProt Related Accession: | P05230 |
| Molecular Weight: | 6,698 Da |
| NCBI Full Name: | Fibroblast growth factor 1 |
| NCBI Synonym Full Names: | fibroblast growth factor 1 (acidic) |
| NCBI Official Symbol: | FGF1 |
| NCBI Official Synonym Symbols: | AFGF; ECGF; FGFA; ECGFA; ECGFB; FGF-1; HBGF1; HBGF-1; GLIO703; ECGF-beta; FGF-alpha |
| NCBI Protein Information: | fibroblast growth factor 1; heparin-binding growth factor 1; beta-endothelial cell growth factor; endothelial cell growth factor, beta; endothelial cell growth factor, alpha |
| UniProt Protein Name: | Fibroblast growth factor 1 |
| UniProt Synonym Protein Names: | Acidic fibroblast growth factor; aFGF; Endothelial cell growth factor; ECGF; Heparin-binding growth factor 1; HBGF-1 |
| Protein Family: | Fibroblast growth factor |
| UniProt Gene Name: | FGF1 |
| UniProt Entry Name: | FGF1_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 | 52264 53020 | 52642 | 52616 |
| 400 | 20049 23419 | 21734 | 21708 |
| 200 | 10307 9109 | 9708 | 9682 |
| 100 | 4534 4570 | 4552 | 4526 |
| 50 | 2240 2136 | 2188 | 2162 |
| 25 | 1144 976 | 1060 | 1034 |
| 12.50 | 501 517 | 509 | 483 |
| 0 | 26 26 | 26 | -- |
Precision
Intra-assay Precision (Precision within an assay): 3 samples with low, mid range and high level Human AFGF/FGF1 were tested 20 times on one plate, respectively.
Inter-assay Precision (Precision between assays): 3 samples with low, mid range and high level Human AFGF/FGF1 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) | 42.84 | 82.30 | 327.26 | 40.45 | 89.02 | 353.66 |
| Standard deviation | 4.36 | 6.32 | 28.01 | 4.91 | 8.46 | 37.84 |
| C V (%) | 10.18 | 7.68 | 8.56 | 12.14 | 9.50 | 10.70 |
Recovery
The recovery of Human AFGF/FGF1 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) | 95-111 | 103 |
| EDTA plasma (n=5) | 94-107 | 101 |
| Cell culture media (n=5) | 88-100 | 93 |
Linearity
Samples were spiked with high concentrations of Human AFGF/FGF1 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 (%) | 85-96 | 90-104 | 91-104 |
| Average (%) | 90 | 95 | 97 | |
| 1:4 | Range (%) | 99-115 | 93-103 | 89-100 |
| Average (%) | 106 | 98 | 94 | |
| 1:8 | Range (%) | 88-102 | 92-105 | 101-118 |
| Average (%) | 94 | 97 | 108 | |
| 1:16 | Range (%) | 93-106 | 91-105 | 87-100 |
| Average (%) | 99 | 97 | 93 |
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.
Related Products
Human FGF1 / FGF acidic ELISA Kit
Bovine Heparin-binding growth factor 1 (FGF1) ELISA Kit
