Human IL-17 ELISA Assay

$430.00

The Human IL-17 ELISA Assay (enzyme-linked immunoassay kit) is intended for the quantitative determination of human Interleukin 17 (IL-17) concentrations in cell culture supernates, serum, and plasma. The Human Interleukin 17 (IL-17) ELISA Assay Kit is for research use only and not to be used in diagnostic procedures.

SKU: L1731-K01 Categories: , ,

Human IL-17 ELISA Assay

The Human IL-17 ELISA Assay is For Research Use Only

Size: 1×96 wells
Sensitivity: 7 pg/mL
Dynamic Range: 15.65 – 500 pg/ml
Incubation Time: 3.5 hours
Sample Type: Serum, Plasma, Cell Culture
Sample Size: 100 µl
Alternative Names: Interleukin 17, CTLA-8


Assay Background

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 Herpes virus saimiri. There is at least some species specificity for in vitro action on bone-marrow stromal cells (3).

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-¿B-mediated mechanism (5). 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 (5) and upregulates nitric oxide (NO) production in osteoarthritic cartilage.


Related Products

Human IL-15 ELISA Assay Kit
Human IL-21 ELISA Assay Kit
Human IL-22 ELISA Assay Kit

Additional Information

Assay Principle


The Eagle Biosciences Human Interleukin 17 (IL-17) ELISA Assay Kit 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 colored 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.

Assay Procedure


  1. Prepare all reagents and working standards as directed in the previous sections.
  2. Determine the number of microwell strips required to test the desired number of samples plus appropriate number of wells needed for running blanks and standards. Remove extra microwell strips from holder and store in foil bag with the desiccant provided at 2-8°C sealed tightly.
  3. Add 100 µL of Standard, control, or sample, per well. Cover with the adhesive strip provided. Incubate for 1.5 hours at 37C.
  4. >Aspirate each well and wash, repeating the process three times for a total of four washes. Wash by filling each well with Wash Buffer (350 µL) using a squirt bottle, manifold dispenser or auto-washer. Complete removal of liquid at each step is essential to good performance. After the last wash, remove any remaining Wash Buffer by aspirating or decanting. Invert the plate and blot it against clean paper towels.
  5. Add 100 µL of the working solution of Biotin-Conjugate to each well. Cover with a new adhesive strip and incubate 1 hour at 37C
  6. Repeat the aspiration/wash.
  7. Add 100 µL of the working solution of Streptavidin-HRP to each well. Cover with a new adhesive strip and incubate for 30 minutes at 37C.  Avoid placing the plate in direct light.
  8. Repeat the aspiration/wash.
  9. Add 100 µL of Substrate Solution to each well. Incubate for 10-20 minutes at 37C.   Avoid placing the plate in direct light.
  10. Add 100 µLof Stop Solution to each well. Gently tap the plate to ensure thorough mixing.
  11. Determine the optical density of each well immediately, using a microplate reader set to 450 nm. (optionally 630nm as the reference wave length;610-650nm is acceptable).

Manual

Product Manual


Publications

References


  1. Rouvier, E. et al. (1993) J. Immunol. 150:5445.
  2. Yao, Z. et al. (1995) J. Immunol. 155:5483.
  3. Kennedy, J. et al. (1996) J. Interferon Cytokine Res. 16:611.
  4. Fossiez, F. et al. (1996) J. Exp. Med. 183:2593.
  5. Yao, Z. et al. (1995) Immunity 3:811.
  6. Ikebuchi, K. et al. (1987) Proc. Natl. Acad. Sci. USA 84:9035.
  7. Berliner, N. et al. (1995) Blood 85:799.
  8. Roberts, A.W. and D. Metcalf (1994) Exp. Hematol. 22:1156.
  9. Broxmeyer, H.E. (1996) J. Exp. Med. 183:2411.
  10. Attur, M.G. et al. (1997) Arthritis Rheum. 40:1050.