Human IL-12 p70 Quantikine HS ELISA Kit

  (6 citations)
(1 Review)
    
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Assay Procedure
Citations (6)
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  • 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.
Control Available
QC41 , Quantikine Immunoassay Control Group 10 - Please Inquire
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 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 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.
 IL-12 p70/IL-12A [HRP]
Product Datasheets

Certificate of Analysis Lookup

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Preparation and Storage
  • 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
Related Research Areas
Assay Procedure
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.
  1.   Prepare all reagents, standard dilutions, and samples as directed in the product insert.
  2.   Remove excess microplate strips from the plate frame, return them to the foil pouch containing the desiccant pack, and reseal.

  3. 50 µL Assay Diluent
  4.   Add 50 µL of the appropriate Assay Diluent to each well.

  5. 200 µL Standard, Control, or Sample
  6.   Add 200 µL of Standard, Control, or sample to each well. Cover with a plate sealer, and incubate at room temperature for 3 hours.
  7.   Aspirate each well and wash, repeating the process 5 times for a total of 6 washes.

  8. 200 µL Conjugate
  9.   Add 200 µL of Conjugate to each well. Cover with a new plate sealer, and incubate at room temperature for 2 hours.
  10.   Aspirate and wash 6 times.

  11. 50 µL Substrate Solution
  12.   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.

  13. 50 µL Amplifier Solution
  14.   Add 50 µL Amplifier Solution to each well. Cover with a new plate sealer, and incubate at room temperature for 30 minutes.

  15. 50 µL Stop Solution
  16. 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.
Citations:

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.

6 Citations: Showing 1 - 6
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Species
Sample Type
  1. 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;0(0):1-14.
    Species: Human
    Sample Type: Serum
  2. 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;2016(0):4685976.
    Species: Human
    Sample Type: Serum
  3. 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;7(0):31.
    Species: Human
    Sample Type: Plasma
  4. 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;12(2):376-82.
    Species: Human
    Sample Type: Plasma
  5. 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;176(8):5078-83.
    Species: Human
    Sample Type: Plasma
  6. 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, 2005;112(6):797-803.
    Species: Human
    Sample Type: Serum

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Average Rating: 5 (Based on 1 review)

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