Human IL-12 p70 Quantikine ELISA Kit
R&D Systems, part of Bio-Techne | Catalog # D1200
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)
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
Reactivity
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.
ND=Non-detectable
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.
| Condition | NC-37 cells | PBMCs |
| Stimulated | 8.3 pg/mL | 33 pg/mL |
| Unstimulated | ND | ND |
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, 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.
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 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
Gene Symbol
IL12A
Additional IL-12 p70 Products
Product Documents for Human IL-12 p70 Quantikine ELISA Kit
Product Specific Notices for Human IL-12 p70 Quantikine ELISA Kit
For research use only
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⚠ 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.