Natural and recombinant phosphorylated and non-phosphorylated human free IGFBP-1. This assay does not detect IGFBP-1 in complex with IGF-I or IGF-II.
< 0.5% cross-reactivity observed with available related molecules.< 50% cross-species reactivity observed with species tested.
No significant interference observed with available related molecules.
QC204 , Quantikine Immunoassay Control Set 851 Human Free IGFBP-1 -
The Quantikine Human Free IGFBP-1 Immunoassay is a 4.5 hour solid-phase ELISA designed to measure human free IGFBP-1 in cell culture supernates, cell lysates, serum, plasma, urine, and human milk. It contains NS0-expressed recombinant human IGFBP-1 and has been shown to accurately quantitate the recombinant factor. Results obtained using natural human IGFBP-1 showed linear curves that were parallel to the standard curves obtained using the Quantikine kit standards. These results indicate that this kit can be used to determine relative mass values for naturally occurring human free IGFBP-1.
Intra-Assay Precision (Precision within an assay) Three samples of known concentration were tested twenty times on one plate to assess intra-assay precision
Inter-Assay Precision (Precision between assays) Three samples of known concentration were tested in twenty separate assays to assess inter-assay precision. Assays were performed by at least three technicians using two lots of components
The recovery of human Free IGFBP-1 spiked to levels throughout the range of the assay in various matrices was evaluated.
Average % Recovery
Cell Culture Media (n=4)
Cell Lysates (n=4)
EDTA Plasma (n=4)
Heparin Plasma (n=4)
Human Milk (n=4)
To assess the linearity of the assay, samples containing and/or spiked with high concentrations of human Free IGFBP-1 were diluted with 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.
The superfamily of insulin-like growth factor (IGF) binding proteins include the six high-affinity IGF binding proteins (IGFBP) and at least four additional low-affinity binding proteins referred to as IGFBP related proteins (IGFBP-rP). All IGFBP superfamily members are cysteine-rich proteins with conserved cysteine residues, which are clustered in the amino- and carboxy-terminal thirds of the molecule. IGFBPs modulate the biological activities of IGF proteins. Some IGFBPs may also have intrinsic bioactivity that is independent of their ability to bind IGF proteins. Post-translational modifications of IGFBP, including glycosylation, phosphorylation and proteolysis, have been shown to modify the affinities of the binding proteins to IGF. ALS (Acid Labile Subunit) is a liver-derived protein that exists in a ternary complex with Insulin-like Growth Factor (IGF)-binding Protein-3 (IGFBP-3) or IGFBP-5, and either IGF-I or IGF-II. ALS increases the half-life of IGF/IGFBP complexes in circulation.
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.
50 µL Assay Diluent
Add 50 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 2-8 °C 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. Cover with a new plate sealer, and incubate at room temperature for 1 hour.
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.
R&D Systems personnel manually curate a database that contains references using R&D Systems products.
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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