Human IL-17 ELISA Kit

Principle of the Assay

This assay employs the quantitative sandwich enzyme immunoassay technique. A monoclonal antibody specific for IL-17 has been pre-coated onto a microplate. Standards and samples are pipetted into the wells and any IL-17 present is bound by the immobilized antibody. Following incubation unbound samples are removed during a wash step, and then a detection antibody specific for IL-17 is added to the wells and binds to the combination of capture antibody- IL-17 in sample. Following a wash to remove any unbound combination, and enzyme conjugate is added to the wells. Following incubation and wash steps a substrate is added. A coloured product is formed in proportion to the amount of IL-17 present in the sample. The reaction is terminated by addition of acid and absorbance is measured at 450nm. A standard curve is prepared from seven IL-17 standard dilutions and IL-17 sample concentration determined

For use with: Serum, Plasma, and Cell culture supernatants.

For Research Use Only. Not for use in diagnostic procedures.

Target Information

IL-17, originally identified as mouse cytotoxic T lymphocyte-associated antigen-8 (CTLA-8), is produced by activated T lymphocytes, primarily by memory T cells. IL-17 appears to mediate communication between the immune system and the hematopoietic system.

IL-17 is a disulfide-linked homodimer. Each polypeptide has 155 amino acid (aa) residues (predicted mass = 17.5 kDa), including a 19 aa residue hydrophobic leader sequence. There are six cysteines plus one potential N-linked glycosylation site, which is variably glycosylated, at least with recombinant proteins. The aa sequence of human IL-17 is 63% and 58% identical to mouse and rat IL-17 and 72% identical to the thirteenth ORF of Herpesvirus saimiri. There is at least some species specificity for in vitro action on bone-marrow stromal cells.

IL-17 mediation of T cell communication with the hematopoietic system is suggested by two observations. T cell-derived IL-17 induces fibroblasts to produce IL-6, IL-8, ICAM-1 and G-CSF, apparently by an NF-ЌB-mediated mechanism. IL-6 in turn promotes development of granulocyte/macrophage colonies, and G-CSF directs development of neutrophils. IL-17 also enhances proliferation of partially activated T cells and upregulates nitric oxide (NO) production in osteoarthritic cartilage.

GENE ID	3605
SWISS PROT	Q16552
SYNONYMS	IL17; CTLA8; IL-17; CTLA-8; IL-17A

Materials Supplied

Kit Components	96 Wells Quantity/Size
Aluminium pouches with a Microwell Plate coated with monoclonal antibody to human IL-17 (8﹡12)	1 plate
Human IL-17 Standard lyophilized, 1000 pg/ml upon reconstitution	2 vials
Concentrated Biotin-Conjugate anti-human IL-17 monoclonal antibody	2 vials
Streptavidin-HRP solution	2 vials
Standard /sample Diluent	1 bottle
Biotin-Conjugate antibody Diluent	1 bottle
Streptavidin-HRP Diluent	1 bottle
Wash Buffer Concentrate 20x (PBS with 1% Tween-20)	1 bottle
Substrate Solution	1 vial
Stop Solution	1 vial
Adhesive Films	4 pieces
Product data sheet	1 copy

Storage

Storage

Store at 2 - 8°C

Performance Characteristics

REPEATABILITY	The coefficient of variation of both intra-assay and inter-assay were less than 10%.
SENSITIVITY	The minimum detectable dose was 7pg/mL.
ASSAY RANGE	15.6–1,000pg/mL
SPECIFICITY	This assay recognizes both natural and recombinant human IL-17. The factors listed below were prepared at 50ng/ml in Standard /sample Diluent and assayed for cross-reactivity and no significant cross-reactivity or interference was observed.