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.
Mouse IL-12 p40 NonAllele-specific Quantikine ELISA
R&D Systems | Catalog # MP400
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Key Product Details
Assay Length
4.5 hours
Sample Type & Volume Required Per Well
Cell Culture Supernates (50 µL), Serum (25 µL), EDTA Plasma (25 µL), Heparin Plasma (25 µL)
Sensitivity
2.7 pg/mL
Assay Range
11.7-750 pg/mL (Cell Culture Supernates, Serum, EDTA Plasma, Heparin Plasma)
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Product Summary for Mouse IL-12 p40 NonAllele-specific Quantikine ELISA
The Quantikine Mouse IL-12/IL-23 p40 Immunoassay is a 4.5 hour solid phase ELISA designed to measure mouse p40 subunit in IL-12 p70, IL-23, p40 homodimer, and p40 monomer in cell culture supernates, mouse serum, and plasma. It contains recombinant mouse IL-12/23 p40 homodimer and antibodies raised against the p40 subunit of recombinant mouse IL-12 p70. This immunoassay has been shown to accurately quantitate the recombinant mouse IL-12/IL-23 p40. Results obtained using natural mouse IL-12/IL-23 p40 showed dose-response curves that were parallel to the standard curves obtained using the recombinant Quantikine kit standards. These results indicate that this kit can be used to determine relative mass values for natural mouse 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
Mouse
Specificity
Natural and recombinant mouse IL-12/IL-23 p40
Cross-reactivity
Cross-reactivity observed with 1 or more 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, EDTA Plasma, Heparin Plasma, Serum
| Intra-Assay Precision | Inter-Assay Precision | |||||
|---|---|---|---|---|---|---|
| Sample | 1 | 2 | 3 | 1 | 2 | 3 |
| n | 20 | 20 | 20 | 26 | 21 | 27 |
| Mean (pg/mL) | 33.3 | 56.4 | 397 | 36.5 | 62.7 | 405 |
| Standard Deviation | 2.3 | 3.0 | 17.2 | 3.8 | 6.5 | 33.7 |
| CV% | 6.9 | 5.3 | 4.3 | 10.4 | 10.4 | 8.3 |
Recovery for Mouse IL-12 p40 NonAllele-specific Quantikine ELISA
The recovery of mouse 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 | 98-108 |
| EDTA Plasma (n=4) | 99 | 81-112 |
| Heparin Plasma (n=4) | 95 | 80-109 |
| Serum (n=4) | 105 | 86-117 |
Linearity
To assess the linearity of the assay, four or more samples containing and/or spiked with various concentrations of mouse IL-12/IL-23 p40 in each matrix were diluted with Calibrator Diluent and then assayed.
Scientific Data Images for Mouse IL-12 p40 NonAllele-specific Quantikine ELISA
Mouse IL-12/IL-23 p40 Non Allele-specific ELISA 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 Mouse IL-12 p40 NonAllele-specific Quantikine ELISA
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 Mouse IL-12 p40 NonAllele-specific Quantikine ELISA
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 Mouse IL-12 p40 NonAllele-specific Quantikine ELISA
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Protocols
View specific protocols for Mouse IL-12 p40 NonAllele-specific Quantikine ELISA (MP400):
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 50 µL of Standard, Control, or sample to each well. Cover with a plate sealer, and incubate at room temperature for 2 hours.
- Aspirate each well and wash, repeating the process 4 times for a total of 5 washes.
- Add 100 µL of Conjugate to each well. Cover with a new plate sealer, and incubate at room temperature for 2 hours.
- Aspirate and wash 5 times.
- Add 100 µL Substrate Solution to each well. Incubate at room temperature for 30 minutes. PROTECT FROM LIGHT.
- Add 100 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
50 µL Standard, Control, or Sample
100 µL Conjugate
100 µL Substrate Solution
100 µL Stop Solution
Find general support by application which include: protocols, troubleshooting, illustrated assays, videos and webinars.
- ELISA Sample Preparation & Collection Guide
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FAQs for Mouse IL-12 p40 NonAllele-specific Quantikine ELISA
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Q: What is the difference between the Mouse IL-12 p40 Allele-specific (M1240) and Non Allele-specific(MP400) Quantikine ELISA kits?
A: Polymorphisms exist in the mouse IL-12/IL-23 p40 sequence. The monoclonal capture antibody used in M1240 is allele-specific. It recognizes the IL-12/IL-23 p40 variant with methionine at residue 169 (Accession # P43432) in mouse strains A.SW, BALB/c, C3H, DBA/1, C57BL/6, NOR, and 129/J. The MP400 kit recognizes variant and non-variant mouse IL-12/IL-23 p40.
