Biotinylation reagents amine-reactive

Amine-reactive biotinylation reagents contain reactive groups off biotin s valeric acid side chain that are able to form covalent bonds with primary amines in proteins and other molecules. Two basic types are commonly available N-hydroxysuccinimide (NHS) esters and carboxylates. NHS esters spontaneously react with amines to form amide linkages (Chapter 2, Section 1.4),... 

Figure 18.15 NHS-chromogenic-PEG3-biotin contains an amine-reactive NHS ester that can be used to label biomolecules through an amide linkage. The chromogenic bis-aryl hydrazone group within the spacer arm of the reagent allows the degree of biotinylation to be quantified by measuring its absorbance at 354 nm. The compound also contains a hydrophilic PEG spacer, which provides greater water solubility.

Perhaps the most common method of DNA biotinylation is through enzymatic incorporation with the use of a biotin-labeled deoxynucleoside triphosphate. First reported by Langer et al. and Leary et al. in 1981, the procedure is probably the most popular nonradioactive labeling technique reported for oligonucleotide probes. Although biotinylated derivatives of dCTP and dATP are reported in the literature, by far the most frequently employed derivative is biotin—dUTP prepared from the reaction of an amine-modified dUTP with an amine-reactive biotinylation reagent, such as NHS-LC—biotin (Chapter 8, Section 3.1). 

Commonly used biotinylating reagents are usually amine reactive. For biotinylation of your protein, do not dissolve your protein sample in amine-containing buffers, such as TBS, TBS-T, or PBS-T. 

The first class of luminescent biotinylation reagents, derived from rhenium(I) polypyridine complexes [Re(NAN)(CO)3(py-biotin-NCS)]+ (24), have been reported by Lo and co-workers [62], To investigate the amine-specific reactivity... 

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