Cross-reactivity observed with 1 or more available related molecules.< 50% cross-species reactivity observed with species tested.
No significant interference observed with available related molecules.
The Quantikine Mouse IL-12/IL-23 p40 Immunoassay is a 4.5 hour solid phase ELISA designed to measure mouse p40 subunit in IL-12 p70, IL-23, p40 homodimer, and p40 monomer in cell culture supernates, mouse serum, and plasma. It contains recombinant mouse IL-12/23 p40 homodimer and antibodies raised against the p40 subunit of recombinant mouse IL-12 p70. This immunoassay has been shown to accurately quantitate the recombinant mouse IL-12/IL-23 p40. Results obtained using natural mouse IL-12/IL-23 p40 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 mouse IL-12/IL-23 p40.
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 mouse IL-12/IL-23 p40 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)
Heparin Plasma (n=4)
To assess the linearity of the assay, four or more samples containing and/or spiked with various concentrations of mouse IL-12/IL-23 p40 in each matrix were diluted with Calibrator Diluent and then assayed.
Store the unopened product at 2 - 8 °C. Do not use past expiration date.
Background: IL-12/IL-23 p40
Interleukin 12 (IL-12), also known as natural killer cell stimulatory factor (NKSF) or cytotoxic lymphocyte maturation factor (CLMF), is a heterodimeric pleiotropic cytokine made up of a 40 kDa (p40) subunit and a 35 kDa (p35) subunit. The IL-12 p40 subunit is shared by IL-23, another heterodimeric cytokine that has biological activities similar to, as well as distinct from, IL-12. IL-12 is produced by macrophages and B cells and has been shown to have multiple effects on T cells and natural killer (NK) cells. While mouse IL-12 is active on both human and mouse cells, human IL-12 is not active on mouse cells.
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 room temperature for 2 hours.
Aspirate each well and wash, repeating the process 4 times for a total of 5 washes.
100 µL Conjugate
Add 100 µL of Conjugate to each well. Cover with a new plate sealer, and incubate at room temperature for 2 hours.
Aspirate and wash 5 times.
100 µL Substrate Solution
Add 100 µL Substrate Solution to each well. Incubate at room temperature for 30 minutes. PROTECT FROM LIGHT.
100 µL Stop Solution
Add 100 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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