Human IL-12 p70 Quantikine ELISA Kit

R&D Systems | Catalog # D1200

R&D Systems
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Key Product Details

Assay Length

3.5 hours or 4.5 hours

Sample Type & Volume Required Per Well

Cell Culture Supernates (200 µL), Serum (200 µL), EDTA Plasma (200 µL), Heparin Plasma (200 µL), Citrate Plasma (200 µL), Urine (200 µL)

Sensitivity

5 pg/mL

Assay Range

7.8-500 pg/mL (Cell Culture Supernates, Serum, EDTA Plasma, Heparin Plasma, Citrate Plasma, Urine)
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Product Summary for Human IL-12 p70 Quantikine ELISA Kit

The Quantikine Human IL-12 Immunoassay is a 3.5 or 4.5 hour solid phase ELISA designed to measure IL-12 in cell culture supernates, serum, plasma, and urine. 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.

Product Specifications

Assay Type

Solid Phase Sandwich ELISA

Format

96-well strip plate

Measurement

Quantitative ELISA

Detection Method

Colorimetric - 450nm (TMB)

Conjugate

HRP

Species

Human

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.

Sample Values

Serum/Plasma/Urine - Forty samples from apparently healthy volunteers were evaluated for the presence of human IL-12 p70 in this assay. No medical histories were available for the donors used in this study. All samples measured less than the lowest human IL-12 p70 standard, 7.8 pg/mL.

Cell Culture Supernates: NC-37 Burkitt's lymphoma B lymphoblast cells (1 x 106 cells/mL) were cultured for 3 days with and without 10 ng/mL PMA and 25 ng/mL calcium ionophore in RPMI 1640 media supplemented with 2 mM L-glutamine, 100 μg/mL penicillin, 100 μg/mL streptomycin, and 1% Nutridoma.

Human peripheral blood mononuclear cells (1 x 106 cells/mL) were cultured for 18 hours with and without 0.0075% (wt/vol) SAC (Pansorbin®, EMD Millipore) in RPMI 1640 media supplemented with 1% fetal bovine serum.

ConditionNC-37 cellsPBMCs
Stimulated8.3 pg/mL33 pg/mL
UnstimulatedNDND
ND=Non-detectable

精度

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, Citrate Plasma, EDTA Plasma, Heparin Plasma, Serum

Intra-Assay Precision Inter-Assay Precision
Sample 1 2 3 1 2 3
n 20 20 20 20 20 20
Mean (pg/mL) 21.2 228 447 21.0 126 246
Standard Deviation 0.3 3.5 4.9 1.5 5.6 8.1
CV% 1.4 1.5 1.1 7.1 4.4 3.3

Urine

Intra-Assay Precision Inter-Assay Precision
Sample 1 2 3 1 2 3
n 20 20 20 20 20 20
Mean (pg/mL) 25.7 202 436 19.9 115 223
Standard Deviation 1.6 4.8 10.1 1.9 8.3 13.2
CV% 6.2 2.4 2.3 9.5 7.2 5.9

Recovery for Human IL-12 p70 Quantikine ELISA Kit

The recovery of human 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) 90 81-96
Citrate Plasma (n=5) 90 82-94
EDTA Plasma (n=5) 101 96-109
Heparin Plasma (n=5) 99 89-109
Serum (n=5) 100 89-106
Urine (n=5) 89 85-92

Linearity

To assess the linearity of the assay, five samples were spiked with high concentrations of IL-12 in various matrices and diluted with Calibrator Diluent to produce samples with values within the dynamic range of the assay.

Human IL-12 p70 ELISA Linearity

Scientific Data Images for Human IL-12 p70 Quantikine ELISA Kit

Human IL-12 p70 ELISA Cell Culture Supernate/Serum/Plasma Standard Curve

Human IL-12 p70 ELISA Cell Culture Supernate/Serum/Plasma Standard Curve

Human IL-12 p70 ELISA Urine Standard Curve

Human IL-12 p70 ELISA Urine Standard Curve

Preparation and Storage

Shipping

The product is shipped at ambient temperature. Upon receipt, store it immediately at the temperature recommended below.

Stability & Storage

Store the unopened product at 2 - 8 °C. Do not use past expiration date.

Background: IL-12 p70

IL-12 (Interleukin-12) is a secreted heterodimeric cytokine that contains disulfide-linked p35 and p40 subunits. The p40 subunit is also a subunit of the heterodimeric IL-23. IL-12 is produced by macrophages, dendritic cells, monocytes, Langerhans cells, neutrophils, keratinocytes, microglia, and non-germinal center B cells. It acts  through a receptor complex that contains the ligand binding IL-12 R alpha 1 and the signal transducing IL-12 R alpha 2. IL-12 can enhance cytotoxic activity and induce Interferon-gamma production in NK cells, T cells and dendritic epidermal T cells. In conjunction with IL-23 and IL-27, IL-12 promotes the development of a Th1 immune response.

Long Name

Interleukin 12 p70

Alternate Names

IL12 p70

Entrez Gene IDs

3592 (Human); 16159 (Mouse); 84405 (Rat)

Gene Symbol

IL12A

Additional IL-12 p70 Products

Product Documents for Human IL-12 p70 Quantikine ELISA Kit

Certificate of Analysis

To download a Certificate of Analysis, please enter a lot or batch number in the search box below.

Note: Certificate of Analysis not available for kit components.

Product Specific Notices for Human IL-12 p70 Quantikine ELISA Kit

For research use only

⚠ WARNING: This product can expose you to chemicals including N,N-Dimethylforamide, which is known to the State of California to cause cancer. For more information, go to www.P65Warnings.ca.gov.

Citations for Human IL-12 p70 Quantikine ELISA Kit

Customer Reviews for Human IL-12 p70 Quantikine ELISA Kit (6)

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Showing  1 - 5 的 6 reviews Showing All
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  • Human IL-12 p70 Quantikine ELISA Kit
    Name: Anonymous
    Sample Tested: Blood mononuclear cells (PBMCs)
    Verified Customer | Posted 06/22/2025
    Secretion of IL-12p70 by three human PBMC donors stimulated with plate-bound aCD3 or aCD28 antibodies
    Human IL-12 p70 Quantikine ELISA Kit D1200
  • Human IL-12 p70 Quantikine ELISA Kit
    Name: Anonymous
    Sample Tested: Cell culture supernatant
    Verified Customer | Posted 01/21/2025
    Human IL-12 p70 Quantikine ELISA Kit D1200
  • Human IL-12 p70 Quantikine ELISA Kit
    Name: Anonymous
    Sample Tested: Serum and Plasma
    Verified Customer | Posted 02/25/2023
  • Human IL-12 p70 Quantikine ELISA Kit
    Name: Anonymous
    Sample Tested: Serum-free Cell Culture Supernates
    Verified Customer | Posted 02/25/2023
  • Human IL-12 p70 Quantikine ELISA Kit
    Name: komal patel
    Sample Tested: Serum and Porcine Serum
    Verified Customer | Posted 07/25/2020
    Human IL-12 p70 Quantikine ELISA Kit D1200
  • Human IL-12 p70 Quantikine ELISA Kit
    Name: Anonymous
    Sample Tested: macrophage
    Verified Customer | Posted 05/11/2020

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Protocols

View specific protocols for Human IL-12 p70 Quantikine ELISA Kit (D1200):

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, working standards, 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 of the Appropriate Assay Diluent
  3.   Add 50 µL of the appropriate Assay Diluent to each well.
    4. 200 µL Standard, control, or sample
  4.   Add 200 µL of Standard, control, or sample to each well. Cover with a plate sealer.
  5.   For Cell Culture Supernate, Serum & Plasma Samples:  Incubate at room temperature for 2 hours.
    For Urine Samples:  Incubate at room temperature for 1.5 hours.
  6.   Aspirate each well and wash, repeating the process twice for a total of 3 washes.
    7. 200 µL Conjugate
  7.   Add 200 µL of Conjugate to each well, and cover with a new plate sealer.
  8.   For Cell Culture Supernate, Serum & Plasma Samples:  Incubate at room temperature for 2 hours.
    For Urine Samples:  Incubate at room temperature for 1.5 hours.
  9.   Aspirate and wash 3 times.
    10. 200 µL Substrate Solution
  10. Add 200 µL Substrate Solution to each well. Incubate at room temperature for 20 minutes. PROTECT FROM LIGHT.
    11. 50 µL Stop Solution
  11. Add 50 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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