Immunochemical detection

LaRochelle, W.J., and Froehner, S.C. (1986b) Immunochemical detection of proteins biotinylated on nitrocellulose replicas./. Immunol. Meth. 92, 65-71. 

Van der Schans, G.P., Scheffer, A.G., Mars-Groenendijk, R.H., Fidder, A., Benschop, H.P., and Baan, R.A. (1994). Immunochemical detection of adducts of sulfur mustard to DNA of calf thymus and human white blood cells. Chem. Res. Toxicol., 7, 408-413. 

Nakajima T, Elovaara E, Park SS, et al. 1991. Immunochemical detection of cytochrome P450 isozymes induced in rat liver by -hexanc, 2-hexanone and acetyl acetone. Arch Toxicol 65 542-547. 

Cribb AE, Nuss CE, Alberts DW, et al. Covalent binding of sulfamethoxazole reactive metabolites to human and rat liver subcellular fractions assessed by immunochemical detection. Chem Res Toxicol 1996 9(2) 500-507. 

Immunochemical detection of quinol-thioether-deiived protein adducts. Chem. Res. Toxicol. 11 1283-1290... 

K. J. Miller, A. C. Herman Affinity chromatography with immunochemical detection applied to the... 

Bokken, G. C. A. M., Corbee, R. J., van Knapen, F., and Bergwerff, A. A. (2003). Immunochemical detection of Salmonella group B, D and E using an optical surface plasmon resonance biosensor. FEMS Microbiol. Lett. 222, 75-82. 

Irth, H., Oosterkamp, A. J., Van der Welle, W., Tjaden, U. R., and Van der Greef, J., On-line immunochemical detection in liquid chromatography using fluorescein-labelled antibodies. J. Chro-matogr. 633, 65-72 (1993). 

The membrane is rinsed with H2O after protein transfer (electrotransfer or dot blot) and dipped to methanol for some seconds. Immediately after this (avoid drying), the membrane is agitated in Soln. A for maximum 2 min and destained by several changes of Soln. B. When the membrane is dry, blue bands are visible on a slight blue background. Membranes stained by this method are not suitable for any immunochemical detection. 

Immunochemical Detection of Antigens After Electrotransfer (Immunoblotting)... 

Immunochemical detection is possible in one step, if the detecting antibody carries the signal-forming principle. But also cascades of steps are used to identify the first-bound antibody and by it the antigen immobilized by blotting. Antibodies form these cascades, i.e., if the first bound antibody is from species A, e.g., rabbit, a second antibody from other species, e.g., goat, directed against the primary, is used. 

The first step in immunochemical detection of proteins after electrotransfer is blocking the support with an inert material to inactivate further non-specific binding of protein. The blocking reagent should cover the membranes at those areas where no blotted protein is bound and should not react with any of the reactants of immunochemical detection cascade as indicated by no non-specific staining, i.e., resulting in blank background of the membrane. 

When a more specific detection system is used instead, a rigorous sample cleanup may not be necessary. This is actually the case with most of the microbiological and immunochemical detection systems applied in residue analysis. Owing to the selectivity and sensitivity of their detection principle, homogenization with an aqueous buffer is often tire only treatment required prior to analysis. Moreover, these detection systems are usually independent of the sample size as, in many cases, a single drop of milk or tissue fluid is sufficient to carry out a successful analysis. 

The number of detectors that are sensitive and selective enough to be applied online with LC is limited because the solvents used are not compatible, as in the case of immunochemical detection after reversed- or normal-phase LC. The technology of coupling is still under development and not yet available in a large number of laboratories not specialized in techniques such as LC-MS. Therefore, LC separations are frequently followed by offline detection. Confirmatory analysis of suspected liquid chromatographic peaks can be made possible by coupling liquid chromatography with mass spectrometry. Atmospheric-pressure chemical ionization LC-MS has been employed for the identification of six steroid hormones in bovine tissues (448). 

Nakamura, M., Kindaichi, K., and Kagayama, M. 1991. Immunohistochemical and immunochemical detection of a similar epitope in enamel proteins and monocyte macrophage protein recognized by mouse monoclonal antibody MOMA-2. Arch. Oral Biol. 36 619-622. 

Hock, B. and R. Niessner (1995). Immunochemical Detection of Pesticides and Their Metabolites in the Water Cycle. Weinheim, Germany VCH Publishers, pp. 195. 

Kramer, P.M., B.A. Baumann, and P.G. Stoks (1997). Prototype of a newly developed immunochemical detection system for the determination of pesticide residues in water. Anal. Chim. Acta, 347 187-198. 

Li, K., L. A. Woodward, A.E. Kara, et al. 2000. Immunochemical detection of polycyclic aromatic hydrocarbons and 1-hydroxypyrene in water and sediment samples. Anal. Chim. Acta 419 1-8. 

Nording, M., M. Nichkova, E. Spinnel, et al. 2006. Rapid screening of dioxin-contaminated soil by accelerated solvent extraction/purification followed by immunochemical detection. Anal. Bioanal. Chem. 385 357-366. 

Salvador, J.P., F. Sanchez-Baeza, and M.P. Marco. 2008. Simultaneous immunochemical detection of stanozolol and the main human metabolite, 3 -hydroxy-stanozolol, in urine and serum samples. Anal. Biochem. 376 221-228. 

Uchida, K., Khor, O.T., Oya, T., Osawa, T., Yasuda, Y., and Miyata, T. 1997. Protein modification by a Maillard reaction intermediate methylglyoxal. Immunochemical detection of fluorescent 5-methylimidazolone derivatives in vivo. FEBS Lett 410 313-318. 

Bartolone, J., Doughty, D., and Egelrud, T., A non-invasive approach for assessing corneocyte cohesion immunochemical detection of desmoglein I, J. Invest. Dermatol., 96, 596, 1991. 

Leduc, V., Demeulemester, C., Polack, B., Guizard, C., Le-Guern, L., Peltre, G. 1999. Immunochemical detection of egg-white antigens and allergens in meat products. Allergy 54 464-472. 

D4, Dray, F., Maron, E., Tillson, S. A., and Sela, M., Immunochemical detection of prostaglandins with prostaglandin-coated bacteriophage T4 and by radioimmunoassay. Anal. Biochem. 50, 399-408 (1972). 

G.P. Van der Schans, D. Noort, R.H. Mars-Groenendijk, A. Fidder, L.F. Chau, L.P.A. De Jong and H.P. Benschop, Immunochemical detection of sulfur mustard adducts with keratins in the stratum corneum of human skin, Chem. Res. Toxicol., 15, 21-25 (2003). 

Y2. Yan, L. J., Orr, W. C., and Sohal, R. S., Identification of oxidized proteins based on sodium dodecyl sulfate-polyacrylamide gel electrophoresis, immunochemical detection, isoelectric focusing, and microsequencing. Anal. Biochem. 263, 67-71 (1998). 

Figure 11.8. Detection of NOS-dependent S-nitrosylation in vivo, a Immunochemical detection procedure. A primary antibody directed against nitrosothiol groups was used in combination with an enzyme-linked secondary antibody. The dye released by the enzyme is insoluble and thus will precipitate close to the site of formation, b S-nitrosylationinaorticrings. Samplesweie treated with Phenylephrine (PE), acetylcholine (Ach), and the inhibitor N-methylarginine (L-NAME) as indicated. Brown stain deposits indicate S-nitrosylation. The traces above the histology panels illustrate the contraction and relaxation responses evoked by the dmgs.

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