The Quantikine HS Human IL-12 Immunoassay is a 6.5 hour solid phase ELISA designed to measure IL-12 in cell culture supernates, serum, and plasma. It contains Sf 21-expressed recombinant human IL-12 and antibodies raised against the recombinant factor. This immunoassay has been shown to quantitate recombinant human IL-12 accurately. Results obtained using natural human IL-12 showed linear curves that were parallel to the standard curves obtained using the recombinant kit standards. These results indicate that this kit can be used to determine relative mass values for natural human IL-12.
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, Heparin Plasma
The recovery of IL-12 spiked to three different levels throughout the range of the assay in various matrices was evaluated.
Average % Recovery
Cell Culture Media (n=5)
EDTA Plasma (n=5)
Heparin Plasma (n=5)
To assess the linearity of the assay, five samples were spiked with high concentrations of IL-12 in various matrices and diluted with the appropriate 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.
Background: IL-12 p70
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 the appropriate Assay Diluent to each well.
200 µL Standard, Control, or Sample
Add 200 µL of Standard, Control, or sample to each well. Cover with a plate sealer, and incubate at room temperature for 3 hours.
Aspirate each well and wash, repeating the process 5 times for a total of 6 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.
Aspirate and wash 6 times.
50 µL Substrate Solution
Add 50 µL Substrate Solution to each well. Cover with a new plate sealer, and incubate at room temperature for 1 hour. Do not wash the plate.
50 µL Amplifier Solution
Add 50 µL Amplifier Solution to each well. Cover with a new plate sealer, and incubate at room temperature for 30 minutes.
50 µL Stop Solution
Add 50 µL of Stop Solution to each well. Read at 490 nm within 30 minutes. Set wavelength correction to 650 nm or 690 nm.
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