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Azides of Biotin/Desthiobiotin

Labeling of a molecule (e.g. a protein or nucleic acid) with biotin/desthiobiotin is routinely performed for its subsequent affinity purification via streptavidin agarose or detection with fluorescent or HRP-labeled streptavidin. Due to the extremely high affinity of biotin towards streptavidin (KD = 10-15 M), the biotinylated molecule/streptavidin-interaction is essentially irreversible under physiological conditions. Desthiobiotin however, binds less tightly to streptavidin (KD = 10-11 M) and desthiobiotinylated molecules are therefore easily eluted from the complex in the presence of excess Biotin.

Azides of biotinylation reagents can be used for the labeling of terminal Alkyne- and strained Alkyne (e.g. DBCO)-labeled biomolecules via Cu(I)-catalyzed Alknye-Azide (CUAAC) or Cu(I)-free strain-promoted Alkyne-Azide Click Chemistry (SPAAC) reaction, respectively.

 

A number of (desthio)biotinylation reagents with

  • different solubility characteristics (PEG-linker for increased molecule solubility)
  • optimized streptavidin binding characteristics (different PEG-linker length)
  • different cleavability characteristics (Diazo-linker for removal of the biotin moiety upon purification)
are available to fit specific application & molecule requirements.