Dde Biotin Alkyne
Dde Biotin Alkyne is an azide-reactive cleavable biotin probe that allows for efficient recovery of avidin-bound protein complexes in affinity-based assays. This reagent contains a biotin moiety linked to azide group through a spacer arm containing a hydrazine-cleavable Dde moiety. Under mild conditions (2% aqueous hydrazine), the Dde liner is cleaved, releasing the biotin tag and the small molecular fragment (55.07 Da) left on the labeled protein following the cleavage.
Extraordinary strength of the streptavidin-biotin interaction allows for efficient capturing of even highly dilute targets; however, it makes recovery of proteins from affinity resins challenging. Conventional methods to elute biotinylated proteins from immobilized avidin include the following: (i) denaturation of streptavidin by boiling the resin in a denaturing buffer that may include high concentrations of chaotropic salts, (ii) trypsin digestion of proteins while they are bound to the resin, or (iii) elution of proteins with excess free biotin. These protocols can co-elute contaminant proteins by releasing nonspecifically bound proteins and/or naturally biotinylated proteins concurrently with labeled proteins. In addition, some of these methods can cause elution of high levels of resin-based peptides along with the proteins of interest, resulting in further sample contamination.
Dde Biotin Alkyne probe eliminates a major limitation of the streptavidin-biotin affinity purification. This reagent contains a biotin moiety linked to an azide (N3) through a spacer arm containing a containing a hydrazine-cleavable Dde moiety. Captured biomolecules can be efficiently released under mild conditions (2% aqueous hydrazine solution), and the small molecular fragment (55.07 Da) left on the labeled protein following the cleavage. These features make the cleavable probe especially attractive for use in biomolecular labeling and proteomic studies.
1. Yang Y., et al. (2013). Cleavable Trifunctional Biotin Reagents for Protein Labeling, Capture, and Release. Chem. Commun., 48: 5366-86. [PubMed]
2. Matthew E. G., et al. (2017). Comprehensive Mapping of O-GlcNAc Modification Sites Using a Chemically Cleavable Tag. Mol. Biosyst, 12: 1756–59. [PubMed]
3. Gertsik N., et al. (2017). Mapping the Binding Site of BMS-708163 on y-Secretase with Cleavable Photoprobes. Cell Chemical Biology, 32: 3-8. [PubMed]
Abs/Em Maxima: 346/445 nmExtinction Coefficient: 19,000Spectrally Similar Dyes: Alexa Fluor? 350, CF? 350, DyLight 350, AMCAAZDye? 350 is a blue-fluorescent azide-activated probe that reacts with terminal alkynes via a copper-catalyzed click react…
Abs/Em Maxima: 402/424 nmExtinction Coefficient: 35,000Flow Cytometry Laser Line: 405 nmMicroscopy Laser Line: 405 nmSpectrally Similar Dyes: Alexa Fluor? 405, CF? 405, Cascade Blue?, DyLight? 405AZDye? 405 Azide is a water-soluble, pH-insensiti…
Abs/Em Maxima: 346/445 nmExtinction Coefficient: 19,000Spectrally Similar Dyes: Alexa Fluor? 350, CF? 350, DyLight 350, AMCAAZDye? 350 Alkyne (Alexa Fluor? 350 Alkyne equivalent) is a blue-fluorescent, alkyne-activated probe that reacts with azid…
Abs/Em Maxima: 430/537 nmExtinction Coefficient: 15,000Spectrally Similar Dyes: Alexa Fluor? 430, CF? 430AZDye? 430 Azide is a water-soluble, green-fluorescent azide-activated probe that reacts with terminal alkynes via a copper-catalyzed click re…
Abs/Em Maxima: 346/445 nmExtinction Coefficient: 19,000Spectrally Similar Dyes: Alexa Fluor? 350, CF? 350, DyLight 350, AMCAAZDye? 350 DBCO reacts with azides via a copper-free “click chemistry” reaction to form a stable triazole and does not re…
Abs/Em Maxima: 402/424 nmExtinction Coefficient: 35,000Flow Cytometry Laser Line: 405 nmMicroscopy Laser Line: 405 nmSpectrally Similar Dyes: Alexa Fluor? 405, CF? 405, Cascade Blue?, DyLight? 405AZDye? 405 Alkyne reacts with azides via a copper…
Abs/Em Maxima: 494/517 nmExtinction Coefficient: 73.000Flow Cytometry Laser Line: 488 nmMicroscopy Laser Line: 488 nmSpectrally Similar Dyes: Fluorescein, Alexa Fluor? 488, CF? 488A, DyLight? 488, Atto? 488AZDye? 488 Azide (Alexa Fluor? 488 Azi…
Abs/Em Maxima: 402/424 nmExtinction Coefficient: 35,000Flow Cytometry Laser Line: 405 nmMicroscopy Laser Line: 405 nmSpectrally Similar Dyes: Alexa Fluor? 405, CF? 405, Cascade Blue?, DyLight? 405ZDye? 405 DBCO reacts with azides via a copper-fr…
