Download PDF by Christoph Kessler (auth.), Priv.-Doz. Dr. Christoph Kessler: Nonradioactive Labeling and Detection of Biomolecules

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By Christoph Kessler (auth.), Priv.-Doz. Dr. Christoph Kessler (eds.)

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1986); Chiron Urdea et al. (1988) University Adelaide Li et al. (1987) EMBO Salk Institute Sproat et al. (1987) Chu and Orgel (1988) Sect. 2 Amersharn/Boebringer MannheimlPierceiSigma AmershamIBoebringer MannheimlPierceiSigma Weizman Institute Wilchek and Bayer (1988) Strauss (1984); Bayer et al. (1985) Wilchek et al. (1986) Sect. 2 Weizman Institute Wilcheketal. (1986) Sect. 2 Boehringer Mannheiml Lacey and Grant (1987) LTIlPierce Boehringer Mannheim Haselbeck and Hosei, personal communication Boehringer Mannheim Haselbeck and HOsel, personal communication Boehringer Mannheim Haselbeck and Hosel, personal communication; Miihleggeretal.

3 Chemical Labeling In the case of nucleic acids, chemical derivatization can be performed by a number of alternative reactions (for review see Matthews and Kricka, 1988; Kricka, 1992). , 1986). , 1988). These allylamine-derivatized oligonucleotides react with N-hydroxysuccinimide (NHS) esters coupled with the haptens of interest. , 1989a; 1989b). Finally, 5' end labeling via NH2 or SH groups has also been reported. The introduction of suitable marker groups into oligonucleotides via modified phospho diester linkages is also possible (for references see Kricka, 1992).

The incorporated biotin is detected by the binding pro tein avidin, isolated from egg white, or streptavidin, isolated from protein Streptomyces avidinii bacteria. Each protein has four high-affinity binding sites for biotin; the binding constant is K = 1015 mol- 1 (Chaiet and Wolf, 1964; Greene, 1975). The biotin label has been widely used in a variety of different assays, including both direct and indirect formats, for detection of nucleic acids, proteins, and glycans on blots, in solution, or in situ (for references see Bayer and Wilchek, 1990).

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