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.
Porcine IL-12/IL-23 p40 Quantikine ELISA Kit
R&D Systems | Catalog # P1240
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Key Product Details
Assay Length
4.5 hours
Sample Type & Volume Required Per Well
Cell Culture Supernates (100 µL), Serum (25 µL), EDTA Plasma (25 µL), Heparin Plasma (25 µL)
Sensitivity
18.2 pg/mL
Assay Range
46.9-3000 pg/mL (Cell Culture Supernates, Serum, EDTA Plasma, Heparin Plasma)
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Product Summary for Porcine IL-12/IL-23 p40 Quantikine ELISA Kit
The Quantikine Porcine IL-12/IL-23 p40 Immunoassay is a 4.5 hour solid-phase ELISA designed to measure porcine IL-12/IL-23 p40 in cell culture supernates, serum and plasma. It contains CHO cell-expressed recombinant porcine IL-12 and antibodies raised against the recombinant factor. This immunoassay has been shown to accurately quantitate the recombinant factor. Results obtained using natural porcine IL-12/IL-23 p40 showed linear curves that were parallel to the standard curves obtained using the Quantikine kit standards. These results indicate that this kit can be used to determine relative mass values for naturally occurring porcine IL-12/IL-23 p40.
Product Specifications
Assay Type
Solid Phase Sandwich ELISA
Format
96-well strip plate
Measurement
Quantitative ELISA
Detection Method
Colorimetric - 450nm (TMB)
Conjugate
HRP
Species
Porcine
Specificity
Natural and recombinant porcine IL-12/IL-23 p40
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.
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 | 27 | 27 | 27 |
| Mean (pg/mL) | 89.8 | 362 | 819 | 88.8 | 361 | 795 |
| Standard Deviation | 7.3 | 17.7 | 28.0 | 9.2 | 22.9 | 46.1 |
| CV% | 8.1 | 4.9 | 3.4 | 10.4 | 6.3 | 5.8 |
EDTA Plasma, Heparin Plasma, Serum
| Intra-Assay Precision | Inter-Assay Precision | |||||
|---|---|---|---|---|---|---|
| Sample | 1 | 2 | 3 | 1 | 2 | 3 |
| n | 20 | 20 | 20 | 25 | 25 | 25 |
| Mean (pg/mL) | 115 | 447 | 991 | 110 | 454 | 995 |
| Standard Deviation | 7.4 | 22.7 | 33.0 | 10.1 | 33.4 | 64.3 |
| CV% | 6.4 | 5.1 | 3.3 | 9.2 | 7.4 | 6.5 |
Recovery for Porcine IL-12/IL-23 p40 Quantikine ELISA Kit
The recovery of porcine IL-12/IL-23 p40 spiked to three levels throughout the range of the assay in various matrices was evaluated.
| Sample Type | Average % Recovery | Range % |
|---|---|---|
| Cell Culture Supernates (n=4) | 104 | 97-113 |
| EDTA Plasma (n=4) | 92 | 84-99 |
| Heparin Plasma (n=5) | 94 | 85-105 |
| Serum (n=5) | 99 | 88-109 |
Linearity
To assess the linearity of the assay, samples spiked with or containing porcine IL-12/IL-23 p40 in each matrix were diluted with the appropriate Calibrator Diluent to produce samples with values within the dynamic range of the assay.
Scientific Data Images for Porcine IL-12/IL-23 p40 Quantikine ELISA Kit
Porcine IL-12/IL-23 p40 ELISA Calibrator Diluent RD5Y Standard Curve
Porcine IL-12/IL-23 p40 ELISA Calibrator Diluent RD6-3 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/IL-23 p40
Long Name
Interleukin 12/Interleukin 23 p40
Alternate Names
CLMF2, IL-12B, IL12 p40, IL12B, NKSF2
Entrez Gene IDs
Gene Symbol
IL12B
Additional IL-12/IL-23 p40 Products
Product Documents for Porcine IL-12/IL-23 p40 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 Porcine IL-12/IL-23 p40 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 Porcine IL-12/IL-23 p40 Quantikine ELISA Kit
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Protocols
View specific protocols for Porcine IL-12/IL-23 p40 Quantikine ELISA Kit (P1240):
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.
- Add 50 µL of Assay Diluent to each well.
- Add 100 µL of Standard, Control, or sample to each well. Cover with a plate sealer, and incubate at room temperature for 2 hours on a horizontal orbital microplate shaker.
- Aspirate each well and wash, repeating the process 4 times for a total of 5 washes.
- Add 200 µL of Conjugate to each well. Cover with a new plate sealer, and incubate at room temperature for 2 hours on the shaker.
- Aspirate and wash 5 times.
- Add 120 µL Substrate Solution to each well. Incubate at room temperature for 30 minutes on the benchtop. PROTECT FROM LIGHT.
- Add 120 µL of Stop Solution to each well. Read at 450 nm within 30 minutes. Set wavelength correction to 540 nm or 570 nm.
50 µL Assay Diluent
100 µL Standard, Control, or Sample
200 µL Conjugate
120 µL Substrate Solution
120 µL Stop Solution
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