Recombinant human Insulin and natural human, canine, and porcine Insulin. This assay cross-reacts 0.25% with Human Proinsulin.
< 0.5% cross-reactivity observed with available related molecules.Cross-species reactivity observed with 1 or more species tested.
No significant interference observed with available related molecules.
The Quantikine Human/Canine/Porcine Insulin Immunoassay is a 4.5 hour solid-phase ELISA designed to measure Insulin in cell culture supernates, serum, and plasma. It contains S. cerevisiae-expressed recombinant human Insulin and has been shown to accurately quantitate the recombinant factor. Results obtained using natural Insulin 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 Insulin.
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 Insulin 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 containing and/or spiked with high concentrations of Insulin were diluted with 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.
Insulin and the IGFs are members of the insulin family of molecules. IGF-I and II are structurally homologous to proinsulin, and can be thought of as taking the shape of an exaggerated, inverted letter G. Intrachain disulfide bonds link sections of the G. The single chain insulin propeptide consists of a 30 amino acid B chain (aa 2554), a C-peptide (aa 5589), and a 21 aa A chain (aa 90110). Removal of the C-peptide by proteolysis enables the formation of mature Insulin, a disulfidelinked heterodimer of the A and B chains. Circulating C-peptide levels are elevated in hyperinsulinism, obesity, and type II
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.
Aspirate each well and wash, repeating once more for a total of 2 washes.
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.
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.