Human IL-15 Quantikine ELISA Kit

R&D Systems | Catalog # D1500

R&D Systems
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Key Product Details

Assay Length

4.25 hours or 4.5 hours

Sample Type & Volume Required Per Well

Cell Culture Supernates (50 µL), Serum (50 µL), EDTA Plasma (50 µL), Heparin Plasma (50 µL)

Sensitivity

2 pg/mL

Assay Range

3.9-250 pg/mL (Cell Culture Supernates, Serum, EDTA Plasma, Heparin Plasma)
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Product Summary for Human IL-15 Quantikine ELISA Kit

The Quantikine Human IL-15 Immunoassay is a 4.25 or 4.5 hour solid phase ELISA designed to measure human IL-15 in cell culture supernates, serum, and plasma. It contains E. coli-expressed recombinant human IL-15 and antibodies raised against the recombinant factor. This immunoassay has been shown to accurately quantitate the recombinant factor. Results obtained using natural human IL-15 expressed by HuT-102 cells showed linear 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-15.

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-15

Cross-reactivity

< 0.5% cross-reactivity observed with available related molecules. Cross-species reactivity not tested.

Interference

No significant interference observed with available related molecules.

Sample Values

Serum/Plasma - Thirty-seven serum and plasma samples from apparently healthy volunteers were evaluated for the presence of human IL-15 in this assay. All samples measured less than the lowest IL-15 standard, 3.91 pg/mL. No medical histories were available for the donors used in this study. 

Cell Culture Supernates:
Human peripheral blood mononuclear cells (5 x 106 cells/mL) were cultured in RPMI supplemented with 5% fetal bovine serum, 50 μM beta -mercaptoethanol, 2 mM L-glutamine, 100 units/mL penicillin, and 100 μg/mL streptomycin sulfate. The cells were cultured unstimulated or stimulated with 10 μg/mL PHA for 1 and 5 days. Aliquots of the cell culture supernates were removed and assayed for levels of natural human IL-15. All samples measured less than the lowest IL-15 standard, 3.91 pg/mL. 

HuT 102 human cutaneous T cell lymphocyte cells were cultured in a 1:1 mixture of DMEM with high glucose and Hamm's F12 supplemented with a 150-fold final dilution of Nutridoma NS (Boehringer Mannheim) and a 500-fold dilution of defined lipid concentrate (Gibco). An aliquot of the cell culture supernate was removed, assayed for human IL-15, and measured 153 pg/mL.

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) 7.2 22.9 131 9.2 29.0 154
Standard Deviation 0.3 1.3 5.8 0.7 1.6 10.2
CV% 4.2 5.7 4.4 7.6 5.5 6.6

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) 7.7 24.7 140 9.9 32.8 184
Standard Deviation 0.3 1.3 4.8 0.9 2.7 12.2
CV% 3.9 5.3 3.4 9.1 8.2 6.6

Recovery for Human IL-15 Quantikine ELISA Kit

The recovery of human IL-15 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) 98 93-104
EDTA Plasma (n=5) 99 90-112
Heparin Plasma (n=5) 100 91-107
Serum (n=5) 98 92-109

Linearity

To assess the linearity of the assay, five samples were spiked with high concentrations of IL-15 in various matrices and diluted with the appropriate Calibrator Diluent to produce samples with values within the dynamic range of the assay.\line

Human IL-15 ELISA Linearity

Scientific Data Images for Human IL-15 Quantikine ELISA Kit

Human IL-15 ELISA Cell Culture Supernate Standard Curve

Human IL-15 ELISA Cell Culture Supernate Standard Curve

Human IL-15 ELISA Serum/Plasma Standard Curve

Human IL-15 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-15

IL-15 (Interleukin 15) is a cytokine that induces or enhances the differentiation, maintenance, or activation of multiple T cell subsets including NK, NKT, Th17, Treg, and CD8+ memory cells. It also induces dendritic cell differentiation and inflammatory activation, exhibits anti-tumor activity, and inhibits the deposition of lipid in adipocytes. Complexes of IL-15 with cell surface IL-15 R alpha interact with complexes of IL-2 R beta and the Common gamma Chain on adjacent cells. This transpresentation mechanism enables cells to respond to IL-15 even if they do not express IL-15 R alpha. Ligation of membrane-associated IL-15/IL-15 R alpha complexes also induces reverse signaling that promotes activation of the IL-15/IL-15 R alpha expressing cells. The activity of circulating IL-15 is limited by its association with soluble IL-15 R alpha.

Long Name

Interleukin 15

Alternate Names

IL15

Entrez Gene IDs

3600 (Human); 16168 (Mouse); 25670 (Rat); 102119613 (Cynomolgus Monkey); 493682 (Feline)

Gene Symbol

IL15

Additional IL-15 Products

Product Documents for Human IL-15 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-15 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-15 Quantikine ELISA Kit

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  • Human IL-15 Quantikine ELISA Kit
    Name: Antoni Gruca
    Sample Tested: Cell culture supernatant
    Verified Customer | Posted 01/21/2025
    Human IL-15 Quantikine ELISA Kit D1500
  • Human IL-15 Quantikine ELISA Kit
    Name: Anonymous
    Sample Tested: Serum-free Cell Culture Supernates
    Verified Customer | Posted 02/25/2023
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    Name: Anonymous
    Sample Tested: Serum and Plasma
    Verified Customer | Posted 01/22/2020
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    Sample Tested: Serum and Plasma
    Verified Customer | Posted 01/10/2020

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Protocols

View specific protocols for Human IL-15 Quantikine ELISA Kit (D1500):

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. 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 3 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 1 hour.
    For Cell Culture Supernate Samples: Cover with a new plate sealer, and incubate at room temperature for 45 minutes.
  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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