Human IL-12/IL-23 p40 Quantikine ELISA Kit

R&D Systems | Catalog # DP400

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 (100 µL), Serum (100 µL), EDTA Plasma (100 µL), Heparin Plasma (100 µL), Citrate Plasma (100 µL)

Sensitivity

15 pg/mL

Assay Range

31.2-2000 pg/mL (Cell Culture Supernates, Serum, EDTA Plasma, Heparin Plasma, Citrate Plasma)
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Product Summary for Human IL-12/IL-23 p40 Quantikine ELISA Kit

The Quantikine Human IL-12 p40 Immunoassay is a 3.5 or 4.5 hour solid phase ELISA designed to measure IL-12 p40 in cell culture supernates, serum, and plasma. It contains recombinant human IL-12 p40 and antibodies raised against the recombinant factor. This immunoassay has been shown to accurately quantitate recombinant human IL-12 p40. Results obtained using natural human IL-12 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 human IL-12 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

Human

Specificity

Natural and recombinant human IL-12 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

Intra-Assay Precision Inter-Assay Precision
Sample 1 2 3 1 2 3
n 20 20 20 40 40 40
Mean (pg/mL) 119 290 871 127 315 891
Standard Deviation 5.1 11.4 35.5 11.8 19.8 60.6
CV% 4.3 3.9 4.1 9.3 6.3 6.8

Citrate Plasma, EDTA Plasma, Heparin Plasma, Serum

Intra-Assay Precision Inter-Assay Precision
Sample 1 2 3 1 2 3
n 20 20 20 40 40 40
Mean (pg/mL) 151 380 1138 160 399 1145
Standard Deviation 9.9 27.2 62.3 11.0 22.3 63.0
CV% 6.6 7.2 5.5 6.9 5.6 5.5

Recovery for Human IL-12/IL-23 p40 Quantikine ELISA Kit

The recovery of IL-12 p40 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) 96 85-105
Citrate Plasma (n=5) 100 85-112
EDTA Plasma (n=5) 101 92-113
Heparin Plasma (n=5) 96 89-106
Serum (n=5) 98 86-112

Linearity

To assess the linearity of the assay, five samples were spiked with high concentrations of IL-12 p40 in various matrices and diluted with the appropriate Calibrator Diluent to produce samples with values within the dynamic range of the assay.

Human IL-12/IL-23 p40 Homodimer ELISA Linearity

Scientific Data Images for Human IL-12/IL-23 p40 Quantikine ELISA Kit

Human IL-12/IL-23 p40 Homodimer ELISA Cell Culture Supernate Standard Curve

Human IL-12/IL-23 p40 Homodimer ELISA Cell Culture Supernate Standard Curve

Human IL-12/IL-23 p40 Homodimer ELISA Serum/Plasma Standard Curve

Human IL-12/IL-23 p40 Homodimer ELISA Serum/Plasma 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

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 Human 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 Human 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 Human IL-12/IL-23 p40 Quantikine ELISA Kit

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  • Human IL-12/IL-23 p40 Quantikine ELISA Kit
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    Sample Tested: Macrophage cytokine expression
    Verified Customer | Posted 05/10/2018
    Human IL-12/IL-23 p40 Quantikine ELISA Kit DP400
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Protocols

View specific protocols for Human IL-12/IL-23 p40 Quantikine ELISA Kit (DP400):

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. 100 µL Assay Diluent
  3.   Add 100 µL of Assay Diluent to each well.
    4. 100 µL Standard, Control, or Sample
  4.   Add 100 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 3 times for a total of 4 washes.
    6. 200 µL Conjugate
  6.   Add 200 µL of Conjugate to each well.
  7.   For Serum & Plasma Samples: Cover with a new plate sealer, and incubate at room temperature for 2 hours.
    For Cell Culture Supernate Samples: Cover with a new plate sealer, and incubate at room temperature for 1 hour.
  8.   Aspirate and wash 4 times.
    9. 200 µL Substrate Solution
  9.   Add 200 µL Substrate Solution to each well. Incubate at room temperature for 30 minutes. PROTECT FROM LIGHT.
    10. 50 µL Stop Solution
  10. 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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