Swine IL-4 ELISA Assay

$445.00

The Swine IL-4 ELISA Assay is intended for the quantitative determination of Swine Interleukin 4 (IL-4) concentrations in cell culture supernates, serum, and plasma. The Swine Interleukin 4 (IL-4) ELISA Assay Kit is for research use only and not to be used in diagnostic procedures.

Swine IL-4 ELISA Assay

The Swine IL-4 ELISA Assay is For Research Use Only

Size: 1×96 wells
Sensitivity: 7 pg/mL
Dynamic Range: 15.625 – 500 pg/ml
Incubation Time: 3.5 hours
Sample Type: Serum, Plasma, Cell Culture
Sample Size: 100 µl
Alternative Names: Porcine Interleukin 4, Porcine IL-4, Swine Interleukin 4, B cell Stimulatory Factor 1


Assay Background

Interleukin 4 (IL-4),  also known as B cell stimulatory factor 1, is a monomeric, approximately 13-18 kDa Th2 cytokine that shows pleiotropic effects during immune responses.It is a glycosylated polypeptide that contains three intrachain disulfide bridges and adopts a bundled four αhelix structure. Porcine IL-4 is synthesized with a 24 amino acid (aa) signal sequence. Mature porcine IL-4 shares 78%, 59%, 41%, and 41% aa sequence identity with bovine, human, mouse, and rat IL-4, respectively. Human IL-4 is active on porcine vascular endothelial cells. IL-4 exerts its effects through two receptor complexes. The type I receptor, which is expressed on hematopoietic cells, is a heterodimer of the ligand binding IL-4 Rα and the common γ chain (a shared subunit of the receptors for IL-4, -7, -9, -15, and -21). The type II receptor on nonhematopoietic cells consists of IL-4Rα and IL13Rα1. The type II receptor also transduces IL-13 mediated signals. IL-4 is primarily expressed by Th2-biased CD4+ T cells, mast cells, basophils, and eosinophils. It promotes cell proliferation, survival, and immunoglobulin class switch to IgE in B cells, acquisition of the Th2 phenotype by naïve CD4+ T cells, priming and chemotaxis of mast cells, eosinophils, and basophils, and the proliferation and activation of epithelial cells. IL-4 plays a dominant role in the development of allergic inflammation and asthma.


Related Products

Swine IL-2 ELISA Assay
Swine IL-10 ELISA Assay Kit
Human IL-4 ELISA Assay

Additional Information

Assay Principle


The Eagle Biosciences Swine Interleukin 4 (IL-4) ELISA Assay Kit employs the quantitative sandwich enzyme immunoassay technique. A monoclonal antibody specific for IL-4 has been pre-coated onto a microplate. Standard, control, or sample and the working solution of Biotin-Conjugate are pipetted into the wells. Following incubation and wash steps any IL-4 present is bound by the immobilized antibody and the detection antibody specific for IL-4 is binds to the combination of capture antibody-IL-4 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-4 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-4 standard dilutions and IL-4 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, then add 50 µl of the working solution of Biotin-Conjugate to each well. Cover with the adhesive strip provided and incubate 2 hours at RT. Adequate mixing is very important for good result. Use a mini-vortexer at the lowest frequency.
  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 Streptavidin-HRP to each well. Cover with a new adhesive strip and incubate for 30 minutes at RT Avoid placing the plate in direct light.
  6. Repeat the aspiration/wash.
  7. Add 100 µl of Substrate Solution to each well. Incubate for 10-20 minutes at RT. Avoid placing the plate in direct light.
  8. Add 100 µl of Stop Solution to each well. Gently tap the plate to ensure thorough mixing.
  9. 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)

Typical Standard Curve


swine il4 elisa standard curve

Documents

Product Manual


 

Please note: All documents above are for reference use only and should not be used in place of the documents included with this physical product. If digital copies are needed, please contact us.

Publications

References


  • Benczik, M. and S.L. Gaffen (2004) Immunol. Invest. 33:109.
  • Chomarat, P. and J. Banchereau (1998) Int. Rev. Immunol. 17:1.
  • Bailey, M. et al. (1993) Biochim. Biophys. Acta 1171:328.
  • Redfield, C. et al. (1991) Biochemistry 30:11029.
  • Stocker, C.J. et al. (2000) J. Immunol. 164:3309.
  • Mueller, T.D. et al. (2002) Biochim. Biophys. Acta 1592:237.
  • Nelms, K. et al. (1999) Annu. Rev. Immunol. 17:701.Paludan, S.R. (1998) Scand. J. Immunol. 48:459.
  • Corthay, A. (2006) Scand. J. Immunol. 64:93.
  • Ryan, J.J. et al. (2007) Crit. Rev. Immunol. 27:15.
  • Grone, A. (2002) Vet. Immunol. Immunopathol. 88:1.
  • Rosenberg, H.F. et al. (2007) J. Allergy Clin. Immunol. 119:1303.

Citations


Ren, J;Lu, H;Wen, S;Sun, W;Yan, F;Chen, X;Jing, J;Liu, H;Liu, C;Xue, F;Xiao, P;Xin, S;Jin, N;, Enhanced immune responses in pigs by DNA vaccine coexpressing GP3 and GP5 of European type porcine reproductive and respiratory syndrome virus, Journal of Virol. Methods, Volume 206C, Page 27-37, https://www.sciencedirect.com/science/article/pii/S0166093414002134

Product Citations