HAZA Biotin Azide

Catalog#
Unit Size
Price (USD)
Availability
Qty
1435-1
1 mg
$89.00
In stock
1435-5
5 mg
$239.00
In stock
1435-25
25 mg
$679.00
In stock

HAZA Biotin Azide is a second generation of a popular Diazo Biotin Azide probe. This sodium dithionite (Na2S2O4) cleavable linker is structurally optimized azobenzene scaffold with significantly improved cleavage kinetics and efficiency. The efficiency of selective cleavage of HAZA linker is at least 100-fold higher compared to Diazo linker [1]. Proteins captured onto streptavidin beads with HAZA Biotin Azide probe can be eluded with 5 mM sodium dithionite in 5 minutes with more than 95% efficiency [2]. In comparison, elution of proteins captured with Diazo linker requires 100 mM sodium dithionite or several elutions with 25 mM dithionite [3].

Molecular weight
742.85
Molecular weight left behind
220.23
Chemical composition
C34H46N8O9S
CAS
n/a
Solubility
DMSO, DMF
Appearance
Dark orange solid
Storage conditions
-20°C.
Shipping conditions
Ambient temperature

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.

HAZA Biotin Azide probes help overcome a major limitation of the streptavidin-biotin affinity purification. This reagent contains a biotin moiety linked to an azide moiety through a spacer arm containing a cleavable linker. Captured biomolecules can be efficiently released under mild conditions (5 mM sodium dithionite) and the small (220.23 Da) molecular fragment left on the labeled protein following cleavage.

 

HAZA Biotin Azide is a second generation of a popular Diazo Biotin Azide probe. This sodium dithionite (Na2S2O4) cleavable linker is structurally optimized azobenzene scaffold with significantly improved cleavage kinetics and efficiency. The efficiency of selective cleavage of HAZA linker is at least 100-fold higher compared to Diazo linker [1]. Proteins captured to streptavidin beads with HAZA Biotin Azide probe can be eluded with 5 mM sodium dithionite in 5 minutes with more than 95% efficiency[2]. In comparison, elution of proteins captured with Diazo linker requires 100 mM sodium dithionite or several elutions with 25 mM dithionite [3].

Combination of very efficient elution and mild cleavage conditions makes HAZA Biotin Azide probe especially attractive for use in biomolecular labeling and proteomic studies.

1. Leriche G., et al. (2010). Optimization of the Azobenzene Scaffold for Reductive Cleavage by Dithionite; Development of an Azobenzene Cleavable Linker for Proteomic Applications. Eur. J. Org. Chem. 23: 4360–64. [PubMed]

2. Chuch N.C., et al. (2010). Nondenaturing Chemical Proteomics for Protein Complex Isolation and Identification ChemBioChem.. 11: 2359–61. [PubMed]

3. Steven H. L., et al. (2007). A Mild Chemically Cleavable Linker System for Functional Proteomic Applications.Angew. Chem. Int. Ed.,. 21: 1284-6. [PubMed]

4. Ying-Yu Y., et al. (2011). Identification of lysine acetyltransferase p300 substrates using 4-pentynoyl-coenzyme A and bioorthogonal proteomics.Bioorg. Med. Chem. Lett.,. 21: 4976-79. [PubMed]

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