In vitro ELISA assays / investigation of interstitial liquid (suction blister fluid)

Anti-Pollution Matrix > Methods > Method list
> In vitro ELISA assays / investigation of interstitial liquid (suction blister fluid)

Method

The ELISA (Enzyme-linked Immunosorbent Assay) is used for the quantitative detection of a specific protein in a sample.

Direct, indirect, competitive and sandwich ELISA can be distinguished. In direct ELISA, the antigen bound to the bottom of the microplate is coupled by a specific antibody. The antibody is labeled with an enzyme that allows the detection. Usually, this enzyme is horseradish peroxidase (HRP) or alkaline phosphatase (AP). In indirect ELISA, an unlabeled primary antibody first binds specifically to the antigen to be detected. This primary antibody is then bound by a polyclonal secondary antibody coupled to a detection enzyme.

ELISA assays can be used to analyze markers of inflammation or markers of oxidative stress, e.g. after contact with air pollutants in the interstitial fluid of suction blister samples.

 

Detection of

• 8-isoprostane (lipid peroxidation), carbonylated proteins, interleukines, collagene and MMPs in the suction blister fluid.

 

Suitable for

• Depending on the assay, different samples (cell lysate, cell and tissue supernatants, or interstitial fluid) can be analyzed.

 

References

• M.R. Barr, S.L. Walker,W.Tsang, G.I. Harrison, P. Ettehadi, M.W. Greaves and A.R. Young. Suppressed alloantigen presentation, increased TNF-α, IL-1, IL-1Ra, IL-10, and modulation of TNF-R in UV-irradiated human skin. J Invest Dermatol. 112(5); 692-698 (1999), DOI: 10.1046/j.1523-1747.1999.00570.x
• K. M. Südel, K. Venzke, E. Knußmann-Hartig, I. Moll, F. Stäb, H. Wenck, K.Wittern, G. Gercken and S. Gallinat. Tight control of matrix metalloproteinase-1 activity in human skin. Photochemistry and photobiology. 78(4); 355 – 360 (2003), DOI: 10.1562/0031-8655(2003)078<0355:tcomma>2.0.co;2
• S. Kuhn, R. Wolber, L. Kolbe, O. Schnorr, H. Sies. Solar-simulated radiation induces secretion of IL-6 and production of istoprostanes in human skin in vivo. Arch Dermatol Res. 297; 477-479 (2006), DOI: 10.1007/s00403-006-0648-2