Human IL-2 Quantikine QuicKit ELISA
R&D Systems, part of Bio-Techne | Catalog # QK202
Key Product Details
Assay Length
80 minutes
Sample Type & Volume Required Per Well
Cell Culture Supernates (50 µL), Serum (50 µL), EDTA Plasma (50 µL), Heparin Plasma (50 µL)
Sensitivity
2.01 pg/mL
Assay Range
31.3-2000 pg/mL (Cell Culture Supernates, Serum, EDTA Plasma, Heparin Plasma)
Product Summary for Human IL-2 Quantikine QuicKit ELISA
The Quantikine® QuicKit™ Human IL-2 Immunoassay is a one step, 80-minute solid phase ELISA designed to measure human IL-2 levels in cell culture supernates, serum, and plasma. It contains E.
coli-expressed recombinant human IL-2 and antibodies raised against the recombinant protein. Results obtained for naturally occurring human IL-2 showed linear curves that were parallel to the standard curves obtained using the recombinant QuicKit™ standards. These results indicate that this kit can be used to determine relative mass values for natural IL-2.
Product Specifications
Measurement
Quantitative ELISA
Detection Method
Colorimetric - 450nm (TMB)
Conjugate
HRP
Reactivity
Human
Specificity
Natural and recombinant human IL-2.
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.
Sample Values
Serum/Plasma - Ten serum and plasma samples from apparently healthy volunteers were
evaluated for the presence of human IL-2 in this assay. All samlpes measured less than the
lowest standard, 31.3 pg/mL. No medical histories were available for the donors used in this
study.
Cell Culture Supernates - Human peripheral blood mononuclear (PBMCs) cells were cultured
in RPMI supplemented with 10% fetal bovine serum, 2 mM L-glutamine, 100 U/mL penicillin,
and 100 μg/mL streptomycin sulfate. The cells were cultured unstimulated or stimulated with
10 μg/mL PHA for 24 hours. Aliquots of the cell culture supernates were removed, assayed for
levels of human IL-2, and were undetectable or measured 390 pg/mL, respectively.
CD4+ T cells were isolated from PBMCs and cultured in RPMI supplemented with 10% fetal
bovine serum, 2 mM L-glutamine, 100 U/mL penicillin, and 100 μg/mL streptomycin sulfate.
Cells were stimulated with plate-bound Mouse Anti-human CD3 (Catalog # MAB100) and
soluble Mouse anti-human CD28 (Catalog # MAB342) for 5 days, followed by 10 ng/mL PMA
and 500 ng/mL Calcium Ionomycin for 24 hours. An aliquot of the cell culture supernate was
removed, assayed for human IL-2, and measured 84,350 pg/mL.
Precision
Intra-Assay Precision (Precision within an assay) Two samples of known concentration were tested twenty times on one plate to assess intra-assay precision.
Inter-Assay Precision (Precision between assays) Two samples of known concentration were tested in ten separate assays to assess inter-assay precision. Assays were performed by at least three technicians.
Cell Culture Supernates, EDTA Plasma, Heparin Plasma, Plasma, Serum
| Intra-Assay Precision | Inter-Assay Precision | |||
|---|---|---|---|---|
| Sample | 1 | 2 | 1 | 2 |
| n | 20 | 20 | 10 | 10 |
| Mean (pg/mL) | 217 | 1231 | 213 | 1331 |
| Standard Deviation | 5.70 | 33.1 | 17.1 | 67.7 |
| CV% | 2.6 | 2.7 | 8.0 | 5.1 |
Recovery for Human IL-2 Quantikine QuicKit ELISA
The recovery of human IL-2 spiked to three levels in samples throughout the range of the assay was evaluated.
| Sample Type | Average % Recovery | Range % |
|---|---|---|
| Cell Culture Media (n=4) | 112 | 103-119 |
| EDTA Plasma (n=2) | 85 | 72-96 |
| Heparin Plasma (n=2) | 85 | 74-93 |
| Serum (n=2) | 91 | 73-105 |
Linearity
To assess the linearity of the assay, samples were spiked with high concentrations of human IL-2 in various matrices and diluted with 1X Kit Diluent to produce samples with values within the dynamic range of the assay.
Scientific Data Images for Human IL-2 Quantikine QuicKit ELISA
Human IL-2 ELISA Standard Curve
Human IL-2 QuicKit Spiked Recovery Competitor Comparison
IL-2 is spiked at three known concentrations throughout the range of the assay and run to measure response of the spiked sample matrix. Serum recovery is 103% compared to 113% for the top competitor. EDTA Plasma recovery is 107% compared to 111% for the top competitor. Heparin Plasma recovery is 100% compared to 102% for the top competitor. In spike and recovery experiments, natural samples are spiked with the recombinant target analyte of interest to identify interference caused by sample matrices.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-2
The receptor for IL-2 (IL-2 R) is composed of three subunits, the 55 kDa CD25/IL-2 R alpha chain, the 70 kDa IL-2 R beta chain, and the 65 kDa Common gamma Chain (1, 3). IL-2 first binds to CD25, the binary complex then recruits IL-2 R beta and gammac to form the quaternary signaling complex (1, 14). In addition to IL-2, IL-2 R beta is used by IL-15 in its quaternary signaling complex. gammac also serves as a signaling receptor for IL-4, -7, -9, -15, and -21 (1, 3).
In vitro studies have shown an important role for IL-2 in T cell activation and expansion. In vivo, IL-2 is critical for the development, maintenance and function of regulatory T cells (Treg) which provide protection against autoimmune disease. On the other hand, IL-2 can also promote autoimmune inflammation in target organs through its roles in regulating the expression of T cell trafficking genes, and production of Th2 cytokines. Within the CD8+ T cell subset, IL-2 is essential for optimal primary responses and differentiation into terminal effector cells. IL-2 also promotes the development of activated CD8+ T cells into memory cells. (1).
Long Name
Interleukin 2
Alternate Names
Aldesleukin, IL2, Proleukin, TCGF
Entrez Gene IDs
Gene Symbol
IL2
Additional IL-2 Products
Product Documents for Human IL-2 Quantikine QuicKit ELISA
Product Specific Notices for Human IL-2 Quantikine QuicKit ELISA
For research use only
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