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Human IL-17 Quantikine ELISA Kit

R&D Systems | Catalog # D1700

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Key Product Details

Assay Length

4.5 hours

Sample Type & Volume Required Per Well

Cell Culture Supernates (100 µL), Serum (100 µL), EDTA Plasma (100 µL), Heparin Plasma (100 µL), Citrate Plasma (100 µL)

Sensitivity

15 pg/mL

Assay Range

31.2-2000 pg/mL (Cell Culture Supernates, Serum, EDTA Plasma, Heparin Plasma, Citrate Plasma)
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Product Summary for Human IL-17 Quantikine ELISA Kit

The Quantikine Human IL-17 Immunoassay is a 4.5 hour solid phase ELISA designed to measure human IL-17 in cell culture supernates, serum, and plasma. It contains E. coli-expressed recombinant human IL-17 and antibodies raised against the recombinant factor. This immunoassay has been shown to quantitate recombinant human IL-17 accurately. Results obtained using natural human IL-17 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-17.

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-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.

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) 107 604 1217 76.6 272 549
Standard Deviation 7.8 25.0 63.3 6.5 23.4 43.9
CV% 7.3 4.1 5.2 8.5 8.6 8.0

Citrate Plasma, 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) 131 736 1480 125 408 794
Standard Deviation 5.8 34.6 61.2 9.8 34.3 55.6
CV% 4.4 4.7 4.1 7.8 8.4 7.0

Recovery for Human IL-17 Quantikine ELISA Kit

The recovery of human IL-17 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) 97 88-112
Citrate Plasma (n=5) 105 92-116
EDTA Plasma (n=5) 101 90-110
Heparin Plasma (n=5) 101 93-109
Serum (n=5) 98 90-105

Linearity

To assess the linearity of the assay, five samples spiked with high concentrations of IL-17 were diluted with the appropriate Calibrator Diluent to produce samples with values within the dynamic range of the assay.

Human IL-17/IL-17A ELISA Linearity

Scientific Data Images for Human IL-17 Quantikine ELISA Kit

Human IL-17/IL-17A ELISA Cell Culture Supernate Standard Curve

Human IL-17/IL-17A ELISA Cell Culture Supernate Standard Curve

Human IL-17/IL-17A ELISA Serum/Plasma Standard Curve

Human IL-17/IL-17A 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-17/IL-17A

Human Interleukin 17 (IL-17), also known as IL-17A and CTLA-8, is a 15-20 kDa, variably glycosylated polypeptide that belongs to the IL-17 family of cytokines (1-5). Its alternate name, CTLA-8, originated from rodent studies where an activated hybridoma was created from the fusion of a mouse cytotoxic and a rat T cell lymphoma cell line. The molecule of interest in this study was assumed to have come from the mouse cytotoxic lymphocyte cell (thus the CTL designation), whereas, in fact, it was a rat lymphocyte molecule. Human IL-17/17A is synthesized as a 155 amino acid (aa) precursor that contains a 23 aa signal sequence and a 133 aa mature region that possesses a cysteine-knot fold (4-6). In both human and mouse, there is one conserved N-linked glycosylation site that likely contributes 5 kDa to its native molecular weight. IL-17A forms both a 32-38 kDa disulfide-linked homodimer, and a 40-45 kDa covalent heterodimer with IL-17F (7-9). Most secreted IL-17A is in the form of the IL-17A:F heterodimer, however, the IL-17A:A homodimer is the most bioactive of the two forms (8). Mature human IL-17A is 61%, 74%, and 99% aa identical to mouse, porcine, and chimpanzee IL-17A, respectively (10-12). Mammalian cells known to produce IL-17 are the CD4+ Th17 T cells, Paneth cells, GR1+CD11b+ myeloid suppressor cells, CD27- gamma delta T cells, CD1+NK1.1- iNKT cells, and CD3- CD4+ LTi-like cells (9, 13-17). 
A high affinity receptor for human IL-17 has been reported, and appears to be a heteromultimer of IL-17RA and IL-17RC, likely in a 2:1 ratio (1, 18). IL-17RA is a 130 kDa, type I transmembrane glycoprotein that bears no resemblance to members of the cytokine, TNF or immunoglobulin receptor superfamily (2, 10, 15). IL-17RC is also a type I transmembrane protein, approximately 90-95 kDa in size, that shares less than 30% aa identity with IL-17RA (19, 20). Both receptors are needed for IL-17A and IL-17A:F activity. The two receptors appear to form a functional association following ligand binding to IL-17RA (1, 21, 22). 
IL-17 is best known for its participation in the recruitment and survival of neutrophils (14, 15, 23, 24, 25). Its induction was initially described to be the result of antigen stimulation of dendritic cells, resulting in IL-23 secretion. In a T cell receptor-independent event, IL-23 induces T cell production of IL-17 (14). Once secreted, IL-17 in the bone marrow would seem to induce stromal/fibroblast expression of both G-CSF and stem cell factor (membrane form), an effect that increases polymorphonuclear neutrophils (PMN) differentiation and production. IL-17 may complement this by directly blocking neutrophil apoptosis, promoting greater circulating PMN numbers (23). In the tissues, IL-17 would also seem to promote neutrophil extravasation, principally through its effects on macrophages and endothelial cells (EC). On macrophages, IL-17 induces TNF-alpha, IL-1b and IL-6 production (26). TNF-alpha and IL-1b then act on local ECs to induce G-CSF secretion, an effect that is potentiated by IL-17 (27). IL-17 further contributes to PMN influx by inducing EC CXC chemokine release and NO production, which may increase vascular permeability (14, 28). IL-17 effects are not limited to inflammation. In synovial joints, IL-17 upregulates RANKL expression on osteoblasts. This provides a stimulus for osteoclast formation and subsequent bone resorption (24). In conjunction with IL-4 and CD40L, IL-17A also promotes the generation of IgE secreting cells (29). And in white fat, IL-17A inhibits adipocyte differentiation from preadipocytes, and impairs glucose uptake by mature adipocytes (30).

Long Name

Interleukin 17

Alternate Names

CTLA-8, CTLA8, IL-17A, IL17, IL17A

Entrez Gene IDs

3605 (Human); 16171 (Mouse); 301289 (Rat); 449530 (Porcine); 481837 (Canine); 102119976 (Cynomolgus Monkey)

Gene Symbol

IL17A

Additional IL-17/IL-17A Products

Product Documents for Human IL-17 Quantikine ELISA Kit

Certificate of Analysis

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Note: Certificate of Analysis not available for kit components.

Product Specific Notices for Human IL-17 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-17 Quantikine ELISA Kit

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    Sample Tested: T cells
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    Verified Customer | Posted 10/26/2015
    Standard curve is shown. <br />Buffer: kit dilution buffer<br />Dilution: 1/100
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Protocols

View specific protocols for Human IL-17 Quantikine ELISA Kit (D1700):

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. 100 µL Standard, Control, or Sample
  4.   Add 100 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 twice for a total of 3 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 1 hour.
  7.   Aspirate and wash 3 times.
    8. 200 µL Substrate Solution
  8.   Add 200 µL Substrate Solution to each well. Incubate at room temperature for 30 minutes. PROTECT FROM LIGHT.
    9. 50 µL Stop Solution
  9.   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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