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 Azide 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 (100.07 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.

Specifications

[caption id="attachment_26280" align="alignnone" width="380"] Schematic Workflow[/caption]

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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.

DADPS (dialkoxydiphenylsilane) Biotin Azide probes eliminate 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 DADPS linker. Captured biomolecules can be efficiently released under mild conditions (10% formic acid, 0.5 h) and the small (143 Da) molecular fragment left on the labeled protein following cleavage. These features make the DADPS probe especially attractive for use in biomolecular labeling and proteomic studies.

Specifications

[caption id="attachment_26280" align="alignnone" width="380"] Schematic Workflow[/caption]

The post Click-&-Go® DADPS Protein Enrichment Kit appeared first on VectorLabs.

Click-&-Go^® Protein Capture Kit provides all the necessary reagents to perform capture of alkyne-modified proteins on high capacity streptavidin agarose resin. The kit includes specially formulated components to perform copper-catalyzed click reaction and subsequent capture on high capacity streptavidin agarose. Sufficient material is supplied for 25 enrichments based on the provided protocol. The kit provides alkyne labeled BSA as a positive control.

Specifications

[caption id="attachment_26261" align="alignnone" width="199"] Schematic Workflow[/caption]

The post Click-&-Go® Protein Enrichment Kit appeared first on VectorLabs.