The Quantikine Human KGF Immunoassay is a 5.25 or 5.5 hour solid phase ELISA designed to measure KGF in cell culture supernates, serum, and plasma. It contains E. coli-expressed recombinant human KGF and antibodies raised against the recombinant factor. This immunoassay has been shown to quantitate the recombinant KGF accurately. Results obtained measuring natural human KGF showed dose-response curves that were parallel to the standard curves obtained using the recombinant Quantikine kit standards. These results indicate that this kit can be used to determine relative mass values for natural human KGF.
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.
The recovery of KGF spiked to three different levels throughout the range of the assay in various matrices was evaluated.
Average % Recovery
Cell Culture Media (n=5)
Citrate Plasma (n=5)
EDTA Plasma (n=5)
Heparin Plasma (n=5)
To assess the linearity of the assay, five samples were spiked with high concentrations of KGF in various matrices and diluted with the appropriate Calibrator Diluent to produce samples with values within the dynamic range of the assay.
Preparation and Storage
Store the unopened product at 2 - 8 °C. Do not use past expiration date.
KGF (keratinocyte growth factor), also known as FGF-7, is induced in mesenchymal cells by inflammation or tissue damage. It functions as a paracrine factor during wound healing by inducing the proliferation of nearby epithelial cells. KGF plays a role in kidney morphogenesis, hair follicle development, and keratinocyte differentiation. It signals only through the IIIb splice form of the tyrosine kinase receptor, FGF R2 (IIIb)/KGF R.
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.
100 µL Assay Diluent
Add 100 µL of Assay Diluent to each well.
100 µL Standard, Control, or Sample
Add 100 µL of Standard, control, or sample to each well. Cover with a plate sealer, and incubate at room temperature for 3 hours.
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.
For Serum & Plasma Samples: Cover with a new plate sealer, and incubate at room temperature for 2 hours. For Cell Culture Supernate Samples: Cover with a new plate sealer, and incubate at room temperature for 1.75 hours.
Aspirate and wash 4 times.
200 µL Substrate Solution
Add 200 µL Substrate Solution to each well. Incubate at room temperature for 30 minutes. 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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The data collected includes not only links to publications in PubMed,
but also provides information about sample types, species, and experimental conditions.
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