Human IL-17 Quantikine QuicKit ELISA Summary
Sample Type & Volume Required Per Well
Cell Culture Supernates (50 uL), Serum (50 uL), EDTA Plasma (50 uL), Heparin Plasma (50 uL)
Assay Range
31.3 - 2,000 pg/mL (Cell Culture Supernates, Serum, EDTA Plasma, Heparin Plasma)
Specificity
Natural and recombinant human IL-17.
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 samples from apparently healthy volunteers were evaluated for the
presence of human IL-17 in this assay. All samples 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:
CD4+ T cells were isolated from human PBMCs using the MagCellect™ Human CD4+ T cell
Isolation Kit (R&D Systems® Catalog # MAGH102). CD4+ T cells were then seeded at 5 x 105
/mL
and cultured in RPMI 1640 supplemented with 10% fetal bovine serum, 2 mM L-glutamine,
100 U/mL penicillin, and 100 μg/mL streptomycin sulfate. T cells were left unstimulated in
culture media, or cultured with immobilized Human CD3 epsilon Antibody (R&D Systems,
Catalog # MAB100) coated at 1 μg/mL and 5 μg/mL soluble Human CD28 Antibody
(R&D Systems, Catalog # MAB342) for 5 days. Stimulated cells were then treated with 10 ng/mL
PMA and 500 ng/mL Ionomycin calcium salt (Tocris™, Catalog # 1704) for 24 hours. An aliquot
of cell culture supernate was removed and assayed for human IL-17.
| Condition | (pg/mL) |
| Unstimulated | ND |
| Stimulated | 244 |
ND=Non-Detectable
Product Summary
The Quantikine™ QuicKit Human IL-17 Immunoassay is a one step, 80-minute solid phase ELISA designed to measure human IL-17 levels in cell culture supernates, serum, and plasma. It contains HEK293-expressed recombinant human IL-17 and antibodies raised against the recombinant protein. Results obtained using natural human IL-17 showed linear curves that were parallel to the standard curves obtained using the QuicKit standards. These results indicate that this kit can be used to determine relative mass values for natural human IL-17.
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
Cell Culture Supernates, Serum, EDTA Plasma, Heparin Plasma
|
Intra-Assay Precision |
Inter-Assay Precision |
| Sample |
1 |
2 |
1 |
2 |
| n |
20 |
20 |
10 |
10 |
| Mean (pg/mL) |
197 |
1025 |
201 |
1090 |
| Standard Deviation |
7.09 |
56.4 |
6.41 |
55.2 |
| CV% |
3.6 |
5.5 |
3.2 |
5.1 |
Recovery
The recovery of human IL-17 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 |
106-118 |
| EDTA Plasma (n=2) |
115 |
107-122 |
| Heparin Plasma (n=2) |
110 |
106-116 |
| Serum (n=2) |
83 |
79-86 |
Linearity
To assess the linearity of the assay, samples containing and/or spiked with high concentrations of human IL-17 were diluted with calibrator diluent to produce samples with values within the dynamic range of the assay.
Scientific Data
Human IL-17 Standard Curve QuicKit ELISA
Product Datasheets
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Preparation and Storage
Shipping
The product is shipped at ambient temperature. Upon receipt, store it immediately at the temperature recommended below.
Storage
Store the unopened product at 2 - 8 °C. Do not use past expiration date.
Background: IL-17/IL-17A
The IL-17 family is comprised of at least six proinflammatory cytokines that share a conserved cysteine-knot structure but diverge at the N-terminus. IL-17 family members are glycoproteins secreted as dimers that induce local cytokine production and recruit granulocytes to sites of inflammation. IL-17 is induced by IL-15 and IL-23, mainly in activated CD4+ T cells distinct from Th1 or Th2 cells. IL-17F is the most homologous to IL-17, but is induced only by IL-23 in activated monocytes. IL-17B binds the IL-17B receptor, but not the IL-17 receptor; it is most homologous with IL-17D, which is expressed by resting CD4+ T cells and CD19+ B cells. IL-17E is mainly produced by Th2 cells and recruits eosinophils to lung tissue. IL-17C has a very restricted expression pattern but has been detected in adult prostate and fetal kidney libraries.
Long Name:
Interleukin 17
Entrez Gene IDs:
3605 (Human); 16171 (Mouse); 301289 (Rat); 449530 (Porcine); 481837 (Canine); 102119976 (Cynomolgus Monkey)
Alternate Names:
CTLA8; CTLA-8; CTLA8cytotoxic T-lymphocyte-associated serine esterase 8; Cytotoxic T-lymphocyte-associated antigen 8; IL17; IL-17; IL17A; IL-17A; IL-17Acytotoxic T-lymphocyte-associated protein 8; IL-17CTLA-8; IL17interleukin-17A; interleukin 17 (cytotoxic T-lymphocyte-associated serine esterase 8); interleukin 17A
