The Quantikine Human FGF acidic Immunoassay is a 4.5 hour solid-phase ELISA designed to measure human FGF acidic in cell culture supernates, serum, plasma, and urine. It contains E. coli-expressed recombinant human FGF acidic and has been shown to accurately quantitate the recombinant factor.
Intra-Assay Precision (Precision within an assay) Three samples of known concentration were tested on one plate to assess intra-assay precision
Inter-Assay Precision (Precision between assays) Three samples of known concentration were tested in separate assays to assess inter-assay precision
Cell Culture Supernates, Urine
Serum, EDTA Plasma, Heparin Plasma
The recovery of FGF acidic spiked to levels throughout the range of the assay in various matrices was evaluated.
Average % Recovery
Cell Culture Media (n=4)
EDTA Plasma (n=4)
Heparin Plasma (n=4)
To assess the linearity of the assay, samples spiked with high concentrations of FGF acidic were serially diluted with the appropriate Calibrator Diluent to produce samples with values within the dynamic range of the assay.
Store the unopened product at 2 - 8 °C. Do not use past expiration date.
Background: FGF acidic
FGF acidic, also known as FGF-1, ECGF, and HBGF-1, is a secreted mitogen that stimulates the proliferation of all cells of mesodermal origin and many cells of neuroectodermal, ectodermal, and endodermal origin. It plays a number of roles in development, regeneration, and angiogenesis. FGF acidic is released extracellularly as a disulfide-linked homodimer and is stored in complex with extracellular heparan sulfate. The association of FGF acidic with heparan sulfate is a prerequisite for its subsequent interaction with FGF receptors. Internalized FGF acidic can translocate to the cytosol and to the nucleus. Intracellular FGF acidic functions as a survival factor by inhibiting p53 activity and proapoptotic signaling.
Refer to the product for complete assay procedure.
Bring all reagents and samples to room temperature before use. It is recommended that all samples, standards, and controls be assayed in duplicate.
Prepare all reagents, standard dilutions, and samples as directed in the product insert.
Remove excess microplate strips from the plate frame, return them to the foil pouch containing the desiccant pack, and reseal.
150 µL Assay Diluent
Add 150 µL of Assay Diluent to each well.
50 µL Standard, Control, or Sample
Add 50 µL of Standard, control, or sample to each well. Cover with a plate sealer, and incubate at room temperature for 2 hours on a horizontal orbital microplate shaker.
Aspirate each well and wash, repeating the process 3 times for a total of 4 washes.
200 µL Conjugate
Add 200 µL of Conjugate to each well. Cover with a new plate sealer, and incubate at room temperature for 2 hours on the shaker.
Aspirate and wash 4 times.
200 µL Substrate Solution
Add 200 µL Substrate Solution to each well. Incubate at room temperature for 30 minutes on the benchtop. PROTECT FROM LIGHT.
50 µL Stop Solution
Add 50 µL of Stop Solution to each well. Read at 450 nm within 30 minutes. Set wavelength correction to 540 nm or 570 nm.
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