Human IL-12 p70 Quantikine HS ELISA Kit

R&D Systems | Catalog # HS120

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Key Product Details

Assay Length

6.5 hours

Sample Type & Volume Required Per Well

Cell Culture Supernates (200 µL), Serum (200 µL), EDTA Plasma (200 µL), Heparin Plasma (200 µL)

Sensitivity

0.5 pg/mL

Assay Range

0.625-40 pg/mL (Cell Culture Supernates)
0.781-50 pg/mL (Serum, EDTA Plasma, Heparin Plasma)
View all IL-12 p70 ELISA Kits »

Product Summary for Human IL-12 p70 Quantikine HS ELISA Kit

The Quantikine HS Human IL-12 Immunoassay is a 6.5 hour solid phase ELISA designed to measure IL-12 in cell culture supernates, serum, and plasma. 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 - 450 or 490nm (TMB or AP Substrate)

Conjugate

HRP

Species

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.

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 20 20 20
Mean (pg/mL) 5.32 13.0 25.0 2.86 8.49 29.6
Standard Deviation 0.24 0.71 1.89 0.5 0.85 2.43
CV% 4.5 5.5 7.6 17.5 10.0 8.2

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) 5.26 18.6 35.8 2.86 9.25 36.4
Standard Deviation 0.26 0.77 0.90 0.36 1.39 2.76
CV% 4.9 4.1 2.5 12.6 15.0 7.6

Recovery for Human IL-12 p70 Quantikine HS ELISA Kit

The recovery of 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) 95 85-107
EDTA Plasma (n=5) 102 83-117
Heparin Plasma (n=5) 97 86-111
Serum (n=5) 100 91-115

Linearity

To assess the linearity of the assay, five samples were spiked with high concentrations of IL-12 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 p70 High Sensitivity ELISA Linearity

Scientific Data Images for Human IL-12 p70 Quantikine HS ELISA Kit

Human IL-12 p70 High Sensitivity ELISA Cell Culture Supernate Standard Curve

Human IL-12 p70 High Sensitivity ELISA Cell Culture Supernate Standard Curve

Human IL-12 p70 High Sensitivity ELISA Serum/Plasma Standard Curve

Human IL-12 p70 High Sensitivity 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 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

Entrez Gene IDs

3592 (Human); 16159 (Mouse); 84405 (Rat)

Gene Symbol

IL12A

Additional IL-12 p70 Products

Product Documents for Human IL-12 p70 Quantikine HS 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 p70 Quantikine HS ELISA Kit

For research use only

Citations for Human IL-12 p70 Quantikine HS ELISA Kit

Customer Reviews for Human IL-12 p70 Quantikine HS ELISA Kit (4)

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  • Human IL-12 p70 Quantikine HS ELISA Kit
    Name: Kimberly Steen
    Sample Tested: EDTA Plasma
    Verified Customer | Posted 12/03/2020
    Results were as expected, and the signal was strong. Great standard performance and clear protocol. Reagents were easy to prepare.
    Human IL-12 p70 Quantikine HS ELISA Kit HS120
  • Human IL-12 p70 Quantikine HS ELISA Kit
    Name: Anonymous
    Sample Tested: Serum
    Verified Customer | Posted 12/02/2020
    Assay is easy to use but very long. Takes a total of 6 and a half hours.
    Human IL-12 p70 Quantikine HS ELISA Kit HS120
  • Human IL-12 p70 Quantikine HS ELISA Kit
    Name: Anonymous
    Sample Tested: Serum
    Verified Customer | Posted 12/02/2020
    Though this assay is lengthy, the results are clear and as expected. All reagents are clearly labeled, as always.
    Human IL-12 p70 Quantikine HS ELISA Kit HS120
  • Human IL-12 p70 Quantikine HS ELISA Kit
    Name: Balaji Mahender
    Sample Tested: EDTA Plasma
    Verified Customer | Posted 01/18/2018

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Protocols

View specific protocols for Human IL-12 p70 Quantikine HS ELISA Kit (HS120):

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 the appropriate Assay Diluent to each well.
    4. 200 µL Standard, Control, or Sample
  4.   Add 200 µL of Standard, Control, or sample to each well. Cover with a plate sealer, and incubate at room temperature for 3 hours.
  5.   Aspirate each well and wash, repeating the process 5 times for a total of 6 washes.
    6. 200 µL Conjugate
  6.   Add 200 µL of Conjugate to each well. Cover with a new plate sealer, and incubate at room temperature for 2 hours.
  7.   Aspirate and wash 6 times.
    8. 50 µL Substrate Solution
  8.   Add 50 µL Substrate Solution to each well. Cover with a new plate sealer, and incubate at room temperature for 1 hour. Do not wash the plate.
    9. 50 µL Amplifier Solution
  9.   Add 50 µL Amplifier Solution to each well. Cover with a new plate sealer, and incubate at room temperature for 30 minutes.
    10. 50 µL Stop Solution
  10. Add 50 µL of Stop Solution to each well. Read at 490 nm within 30 minutes. Set wavelength correction to 650 nm or 690 nm.

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FAQs

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