SNAP-Biotin®i s a cell-permeable substrate (BG-Biotin) based on biotin with an amidocaproyl linker. It is suitable for applications such as biotinylation of SNAP-tag® fusion proteins in living cells for detection with streptavidin fluorophore conjugates or labeling in solution for analysis by SDS-PAGE/Western Blot or for capture with streptavidin for binding and interaction studies. This package contains 50 nmol of SNAP-Biotin substrate, sufficient to make 10 ml of a 5 μM solution for the labeling of SNAP-tag fusion proteins in cells.
The SNAP-tag protein labeling system enables the specific, covalent attachment of virtually any molecule to a protein of interest. The SNAP-tag is a protein based on human O6-alkylguanine-DNA alkyltransferase (hAGT). SNAP-tag substrates are fluorophores, biotin or beads conjugated to guanine or chloropyrimidine leaving groups via a benzyl linker. In the labeling reaction, the substituted benzyl group of the substrate is covalently attached to the SNAP-tag.
There are two steps to using this system: subcloning and expression of the protein of interest as a SNAP-tag fusion, and labeling of the fusion with the SNAP-tag substrate of choice. Expression of SNAP-tag fusion proteins is described in the documentation supplied with SNAP-tag plasmids. The labeling of fusion proteins with the SNAP-Biotin substrate is described below.
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