Mouse IL-12 p40 Allele-specific Quantikine ELISA Kt

R&D Systems | Catalog # M1240

R&D Systems
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Key Product Details

Assay Length

4.5 hours

Sample Type & Volume Required Per Well

Cell Culture Supernates (50 µL), Serum (50 µL)

Sensitivity

4 pg/mL

Assay Range

11.7-750 pg/mL (Cell Culture Supernates, Serum)
View all IL-12/IL-23 p40 ELISA Kits »

Product Summary for Mouse IL-12 p40 Allele-specific Quantikine ELISA Kt

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 and mouse serum. It contains recombinant mouse IL-12 p40 homodimer and antibodies raised against the recombinant factor. 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, Serum

Intra-Assay Precision Inter-Assay Precision
Sample 1 2 3 1 2 3
n 20 20 20 20 20 20
Mean (pg/mL) 42.2 161 588 41.9 151 581
Standard Deviation 0.81 1.7 9.6 4.1 10.3 24.0
CV% 1.9 1.1 1.6 9.8 6.8 4.1

Recovery for Mouse IL-12 p40 Allele-specific Quantikine ELISA Kt

The recovery of mouse IL-12/IL-23 p40 spiked to levels in samples throughout the range of the assay was evaluated.

Sample Type Average % Recovery Range %
Cell Culture Supernates (n=11) 100 81-116
Serum (n=7) 104 85-115

Linearity

To assess the linearity of the assay, five 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.

Mouse IL-12/IL-23 p40 Allele-specific ELISA Linearity

Scientific Data Images for Mouse IL-12 p40 Allele-specific Quantikine ELISA Kt

Mouse IL-12/IL-23 p40 Allele-specific ELISA Standard Curve

Mouse IL-12/IL-23 p40 Allele-specific ELISA Standard Curve

Preparation and Storage

Shipping

The product is shipped with polar packs. 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

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/Interleukin 23 p40

Alternate Names

CLMF2, IL-12B, IL12 p40, IL12B, NKSF2

Entrez Gene IDs

3593 (Human); 16160 (Mouse); 64546 (Rat); 397076 (Porcine); 403976 (Canine); 102124858 (Cynomolgus Monkey); 493741 (Feline); 694747 (Rhesus Macaque)

Gene Symbol

IL12B

Additional IL-12/IL-23 p40 Products

Product Documents for Mouse IL-12 p40 Allele-specific Quantikine ELISA Kt

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 Allele-specific Quantikine ELISA Kt

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 Allele-specific Quantikine ELISA Kt

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Protocols

View specific protocols for Mouse IL-12 p40 Allele-specific Quantikine ELISA Kt (M1240):

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
  3.   Add 50 µL of Assay Diluent to each well.
    4. 50 µL Standard, Control, or Sample
  4.   Add 50 µL of Standard, Control, or sample to each well. Cover with a plate sealer, and incubate at room temperature for 2 hours.
  5.   Aspirate each well and wash, repeating the process 4 times for a total of 5 washes.
    6. 100 µL Conjugate
  6.   Add 100 µL of Conjugate to each well. Cover with a new plate sealer, and incubate at room temperature for 2 hours.
  7.   Aspirate and wash 5 times.
    8. 100 µL Substrate Solution
  8.   Add 100 µL Substrate Solution to each well. Incubate at room temperature for 30 minutes. PROTECT FROM LIGHT.
    9. 100 µL Stop Solution
  9.   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.

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FAQs for Mouse IL-12 p40 Allele-specific Quantikine ELISA Kt

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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.
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