Reagent Formatting

Once antibodies and antigens have been developed, they must be incorporated into an assay system for visualization of the primary antibody-antigen reaction. To accomplish this task effectively, an assay format and procedures to visualize the antibody-antigen reaction must be first selected, the necessary reagents must then be prepared, and the final immunochemical method including sample preparation must be optimized for the intended application. 

Common immunochemical assay formats to select from include the 96-well microtiter plates, dipsticks, coated test tubes, and membrane-based flow through devices. If the end-user is a trained technician working in a well-equipped laboratory and needs to detect and tentatively identify, for example, antimicrobial residues in hundreds of meat samples per day, a multiwell or other high-through-put format should be chosen. If, on the other hand, the end user is a quality control inspector at a milk factory who has limited time to find out whether the penicillin residues in the milk waiting to be unloaded exceed a certain level, the same reagents used in the first instance may require a more user-friendly format such as dipstick or membrane-based flow through device. 

Apart from home-made methods, a number of quick, sensitive, and easy-to-perform immunochemical tests are currently available in a kit format. Table 

1 gives an example of the detection limits attainable by some commercially 

Antibacterial CTII test CITE test LacTek EZ-Screen Spot test 

---

Barbier reported (1) in 1899 that a mixture of methyl iodide, a methyl ketone, and magnesium metal in diethyl ether produced a tertiary alcohol. Detailed studies by his student Victor Grignard are documented in his now classical doctoral thesis, presented in 1901. Grignard estabUshed (2) that the reaction observed by Barbier could be separated into three distinct steps Grignard reagent formation, Grignard reaction, and hydrolysis. 

Spontaneous perfluoroalkylatwn ofenamines occurs in the presence of fluor-inated perhalogenoalkanes [743, 744] This condensation is interpreted by a SET process (mechanism analogous to equation 57 with a neutral nucleophile in place of a charged nucleophilic reagent). Formation of chlorodifluoromethylcyclo... 

The possibility of a radical mechanism is supported by the observation of the accelerating effect of molecular oxygen on the cyclopropanation. Miyano et al. discovered that the addition of dioxygen accelerated the formation of the zinc carbenoid in the Furukawa procedure [24a, b]. The rate of this process was monitored by changes in the concentration of ethyl iodide, the by-product of reagent formation. Comparison of the reaction rate in the presence of oxygen with that in the... 

The mechanism of Grignard reagent formation involves free radicals," and... 

Grignard reagent formation amidation of a Lepetit Research Nebuloni and... 

Formation of Grignard reagent formation of Ciba-Geigy Brogli etal. (1981)... 

After quenching with DzO or Bu OD, analysis of the products from the Grignard reagents formed from PhCHXMe (X = Cl, Br, I) in the optically active solvent -(R)-2-methoxypentane leads to the conclusion that Grignard reagent formation occurs on the Mg surface within a solvent cage by a one-electron transfer mechanism.1... 

Presence of reagents Formation of intermediates may he proven hy their reaction with intentionally added reagents, for example, nucleophiles to quench electrogenerated carhenium ions. Characteristic changes are expected, for example, peaks in CV may disappear. 

Correlate reaction the reaction of telluronium salts with carbonyl compounds mediated by organoUthium reagents - formation of secondary alcohols... 

Alkynyl(phenyl)iodonium Tosylates Preparation and Stereospecific Coupling with Vinylcopper Reagents. Formation of Conjugated Enynes P. J. Stang and T. Kitamura, Department of Chemistry University of Utah, Salt Lake City, UT 84112... 

Eor factors controlling Grignard reagent formation, see W. E. Lindsell, in Comprehensive Organometallic Chemistry I (Eds. G. Wilkinson, E. G. S. Stone and G. E. Ebel), Vol. 1, Chap. 3, Pergamon Press, Oxford, 1982, pp. 155-252 and references therein. 

Alkanes can also be prepared from alkyl halides by reduction, directly with Zn and acetic acid (AcOH) (see Section 5.7.14) or via the Grignard reagent formation followed by hydrolytic work-up (see Section 5.7.15). The coupling reaction of alkyl halides with Gilman reagent (R 2CuLi, lithium organocuprates) also produces alkanes (see Section 5.5.2). 

In the cases where their potential decomposition via a /3-hydride elimination pathway is possible, a large excess of the reagent and careful temperature control is necessary to get a high yield of the cyclopropane. The reagent formation is more effective if a diiodo precursor is used instead of dibromides and activated dichlorides, but these latter two have also been successfully converted into suitable reagents. 

Song, J. H. and Sailor, M. J. Functionalization of nanocrystalline porous silicon surfaces with aryllithium reagents Formation of silicon-carbon bonds by cleavage of silicon-silicon bonds. Journal of the American Chemical Society 120, 2376 (1998). 

---