< 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.
The Quantikine Canine IL-6 Immunoassay is a 4.5 hour solid-phase ELISA designed to measure canine IL-6 in cell culture supernates, serum, and plasma. It contains E. coli-expressed recombinant canine IL-6 and antibodies raised against the recombinant factor. This immunoassay has been shown to accurately quantitate the recombinant factor. Results obtained using natural canine IL-6 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 canine IL-6.
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 separate assays to assess inter-assay precision.
The recovery of canine IL-6 spiked to three levels throughout the range of the assay in various matrices was evaluated.
Average % Recovery
Cell Culture Supernates (n=6)
EDTA Plasma (n=4)
Heparin Plasma (n=4)
To assess the linearity of the assay, samples containing or spiked with various concentrations of canine IL-6 in each matrix were diluted with Calibrator Diluent and then assayed.
Preparation and Storage
Store the unopened product at 2 - 8 °C. Do not use past expiration date.
IL-6 (Interleukin-6) is a pleiotropic cytokine that acts in the acute phase reaction, inflammation, hematopoiesis, bone metabolism, and cancer progression. It contributes to chronic inflammation in obesity, insulin resistance, inflammatory bowel disease, arthritis, sepsis, and atherosclerosis. IL-6 signals through a receptor complex of IL-6 R alpha and gp130. gp130 is also a component of the receptors for CLC, CNTF, CT-1, IL-11, IL-27, LIF, and OSM. Soluble forms of IL-6 R alpha are generated by both alternative splicing and proteolytic cleavage. In a mechanism known as trans-signaling, complexes of soluble IL-6 and IL-6 R alpha elicit responses from gp130-expressing cells that lack cell surface IL-6 R alpha.
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.
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 2 hours on an orbital microplate shaker.
Aspirate each well and wash, repeating the process 4 times for a total of 5 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 5 times.
120 µL Substrate Solution
Add 120 µL Substrate Solution to each well. Incubate at room temperature on the benchtop for 30 minutes. PROTECT FROM LIGHT.
120 µL Stop Solution
Add 120 µL of Stop Solution to each well. Gently tap the plate to ensure thorough mixing. Read at 450 nm within 30 minutes. Set wavelength correction to 540 nm or 570 nm.
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