Human IL-15 Quantikine ELISA Kit

Name: Human IL-15 Quantikine ELISA Kit
Brand: R&D Systems
SKU: D1500
Rating: 5 (3 reviews)

R&D Systems, part of Bio-Techne | Catalog # D1500

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Product Datasheet / COA / SDS

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Product Summary
Product Specifications
Sample Values
Precision
Recovery
Linearity
Scientific Data
Preparation & Storage
Background
Product Documents
Specific Notices

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

Reactivity

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

Scientific Data Images for Human IL-15 Quantikine ELISA Kit

Human IL-15 ELISA Cell Culture Supernate 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