< 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 Porcine IL-10 Immunoassay is a 4.5 hour solid phase ELISA designed to measure porcine IL-10 levels in cell culture supernates, porcine serum, and EDTA plasma. It contains E. coli-expressed recombinant porcine IL-10 and antibodies raised against the recombinant protein. Results obtained for naturally occurring porcine IL-10 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 of natural porcine IL-10.
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, Serum, EDTA Plasma
The recovery of porcine IL-10 spiked to three levels throughout the range of the assay in various matrices was evaluated.
Average % Recovery
Cell Culture Supernates (n=4)
EDTA Plasma (n=4)
To assess the linearity of the assay, four samples spiked with high concentrations of porcine IL-10 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.
Interleukin-10 functions as an anti-inflammatory cytokine by inhibiting the expansion and activation of Th1 cells and Th17 cells and by promoting the development of M2 macrophages and regulatory T cells (Treg). Within a tumor microrenvironment, however, IL-10 can inhibit the expansion of both Treg and myeloid-derived suppressor cells (MDSC). IL-10 exerts protective effects including limiting tissue damage in arthritic inflammation and promoting muscle regeneration after injury, but it also contributes to the persistence of viral infections. IL-10 signals through a receptor complex composed of IL-10 R alpha and IL-10 R beta. IL-10 R beta additionally associates with IL-20 R alpha, IL-22 R alpha 1, or IL-28 R alpha to form the receptor complexes for IL-22, IL-26, IL-28, and IL-29.
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 2 hours on a horizontal 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 for 30 minutes on the benchtop. PROTECT FROM LIGHT.
120 µL Stop Solution
Add 120 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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