Summary
Precision
Recovery
Linearity
Data Examples
Product Datasheets
Preparation and Storage
Background
Related Research Areas
Human IL-12 p70 Quantikine HS ELISA Kit Summary
Assay Type
Solid Phase Sandwich ELISA
Format
96-well strip plate
Assay Length
6.5 hours
Sample Type & Volume Required Per Well
Cell Culture Supernates (200 uL), Serum (200 uL), EDTA Plasma (200 uL), Heparin Plasma (200 uL)
Sensitivity
0.5 pg/mL
Assay Range
0.6 - 40 pg/mL (Cell Culture Supernates), 0.8 - 50 pg/mL (Serum, EDTA Plasma, Heparin Plasma)
Specificity
Natural and recombinant human IL-12p70
Cross-reactivity
< 0.5% cross-reactivity observed with available related molecules.< 50% cross-species reactivity observed with species tested.
Interference
No significant interference observed with available related molecules.
Product Summary
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.
Precision
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
| Intra-Assay Precision | Inter-Assay Precision | |||||
|---|---|---|---|---|---|---|
| Sample | 1 | 2 | 3 | 1 | 2 | 3 |
| n | 20 | 20 | 20 | 20 | 20 | 20 |
| Mean (pg/mL) | 5.32 | 13 | 25 | 2.86 | 8.49 | 29.6 |
| Standard Deviation | 0.24 | 0.71 | 1.89 | 0.5 | 0.85 | 2.43 |
| CV% | 4.5 | 5.5 | 7.6 | 17.5 | 10 | 8.2 |
Serum, EDTA Plasma, Heparin Plasma
| Intra-Assay Precision | Inter-Assay Precision | |||||
|---|---|---|---|---|---|---|
| Sample | 1 | 2 | 3 | 1 | 2 | 3 |
| n | 20 | 20 | 20 | 20 | 20 | 20 |
| Mean (pg/mL) | 5.26 | 18.6 | 35.8 | 2.86 | 9.25 | 36.4 |
| Standard Deviation | 0.26 | 0.77 | 0.9 | 0.36 | 1.39 | 2.76 |
| CV% | 4.9 | 4.1 | 2.5 | 12.6 | 15 | 7.6 |
Recovery
The recovery of IL-12 spiked to three different levels throughout the range of the assay in various matrices was evaluated.
| Sample Type | Average % Recovery | Range % |
|---|---|---|
| Cell Culture Media (n=5) | 95 | 85-107 |
| EDTA Plasma (n=5) | 102 | 83-117 |
| Heparin Plasma (n=5) | 97 | 86-111 |
| Serum (n=5) | 100 | 91-115 |
Linearity
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.
Scientific Data
Product Datasheets
Preparation and Storage
Shipping
The product is shipped at ambient temperature. Upon receipt, store it immediately at the temperature recommended below.
Storage
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.
Long Name:
Interleukin 12 p70
Entrez Gene IDs:
3592 (Human); 16159 (Mouse); 84405 (Rat)
Alternate Names:
CLMF p35; Cytotoxic lymphocyte maturation factor 35 kDa subunit; IL12 p70; IL-12 p70; IL-12 subunit p35; IL-12, subunit p35; interleukin 12, p35; interleukin 12A (natural killer cell stimulatory factor 1, cytotoxic lymphocytematuration factor 1, p35); interleukin-12 alpha chain; interleukin-12 subunit alpha; natural killer cell stimulatory factor 1, 35 kD subunit; NF cell stimulatory factor chain 1; NK cell stimulatory factor chain 1; NKSF1; p35
Assay Procedure
Refer to the product 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.
- Add 50 µL of the appropriate Assay Diluent to each well.
- 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.
- 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.
- 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.
- Add 50 µL Amplifier Solution to each well. Cover with a new plate sealer, and incubate at room temperature for 30 minutes.
- 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.
50 µL Assay Diluent
200 µL Standard, Control, or Sample
200 µL Conjugate
50 µL Substrate Solution
50 µL Amplifier Solution
50 µL Stop Solution
Citations for Human IL-12 p70 Quantikine HS ELISA Kit
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.
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Citations: Showing 1 - 8
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The Impact of Metabolic Memory on Immune Profile in Young Patients with Uncomplicated Type 1 Diabetes
Authors: Neubauer-Geryk, J;Wielicka, M;My?liwiec, M;Zorena, K;Bieniaszewski, L;
International journal of molecular sciences
Species: Human
Sample Types: Serum
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Nanoscale, antigen encounter-dependent, IL-12 delivery by CAR T cells plus PD-L1 blockade for cancer treatment
Authors: Z Yang, V Pietrobon, M Bobbin, O Stefanson, J Yang, A Goswami, B Alphson, H Choi, K Magallanes, Q Cai, D Barrett, B Wang, LS Qi, FM Marincola
Journal of Translational Medicine, 2023-02-28;21(1):158.
Species: Human
Sample Types: Cell Culture Supernates
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Effect of combined consumption of Lactobacillus brevis KB290 and ?-carotene on minor diarrhoea-predominant irritable bowel syndrome-like symptoms in healthy subjects: a randomised, double-blind, placebo-controlled, parallel-group trial
Authors: N Fuke, K Aizawa, H Suganuma, T Takagi, Y Naito
Int J Food Sci Nutr, 2017-04-09;0(0):1-14.
Species: Human
Sample Types: Serum
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High Interleukin-12 Levels May Prevent an Increase in the Amount of Fungi in the Gastrointestinal Tract during the First Years of Diabetes Mellitus Type 1
Authors: B Kowalewska, K Zorena, M Szmigiero-, P W??, M My?liwiec
Dis. Markers, 2016-12-29;2016(0):4685976.
Species: Human
Sample Types: Serum
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Randomized comparison of single dose of recombinant human IL-12 versus placebo for restoration of hematopoiesis and improved survival in rhesus monkeys exposed to lethal radiation.
Authors: Gluzman-Poltorak Z, Mendonca S, Vainstein V, Kha H, Basile L
J Hematol Oncol, 2014-04-06;7(0):31.
Species: Human, Primate - Macaca mulatta (Rhesus Macaque)
Sample Types: Plasma
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Human complete Stat-1 deficiency is associated with defective type I and II IFN responses in vitro but immunity to some low virulence viruses in vivo.
Authors: Chapgier A, Wynn RF, Jouanguy E, Filipe-Santos O, Zhang S, Feinberg J, Hawkins K, Casanova JL, Arkwright PD
J. Immunol., 2006-04-15;176(8):5078-83.
Species: Human
Sample Types: Plasma
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Expression of interleukin-18 and caspase-1 in cutaneous T-cell lymphoma.
Authors: Yamanaka K, Clark R, Dowgiert R, Hurwitz D, Shibata M, Rich BE, Hirahara K, Jones DA, Eapen S, Mizutani H, Kupper TS
Clin. Cancer Res., 2006-01-15;12(2):376-82.
Species: Human
Sample Types: Plasma
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Correlations between IL-4, IL-12 levels and CCL2, CCL5 levels in serum and cerebrospinal fluid of multiple sclerosis patients.
Authors: Bartosik-Psujek H, Stelmasiak Z
J Neural Transm, 2004-10-12;112(6):797-803.
Species: Human
Sample Types: Serum
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Reviews for Human IL-12 p70 Quantikine HS ELISA Kit
Average Rating: 4.8 (Based on 4 Reviews)
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Human IL-12 p70 Quantikine HS ELISA Kit
By Kimberly Steen
on 12/03/2020
Sample Tested: EDTA Plasma
Results were as expected, and the signal was strong. Great standard performance and clear protocol. Reagents were easy to prepare.
Human IL-12 p70 Quantikine HS ELISA Kit
By Anonymous
on 12/02/2020
Sample Tested: Serum
Assay is easy to use but very long. Takes a total of 6 and a half hours.
Human IL-12 p70 Quantikine HS ELISA Kit
By Anonymous
on 12/02/2020
Sample Tested: Serum
Though this assay is lengthy, the results are clear and as expected. All reagents are clearly labeled, as always.
Human IL-12 p70 Quantikine HS ELISA Kit
By Balaji Mahender
on 01/18/2018
Sample Tested: EDTA Plasma
