SNAP-Surface® Block (C8 propanoic acid benzylguanine, CBG) is a nonfluorescent compound that blocks the reactivity of the SNAP-tag® in solution or on the surface of living cells. It can be used to generate inactive controls in live cell labeling experiments performed with SNAP-tag fusion proteins. SNAP-Surface Block reacts with the SNAP-tag irreversibly, inactivating it for subsequent labeling steps. This blocker is largely membrane impermeable essentially limiting blocking to cell surface-exposed SNAP-tags.
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: sub-cloning 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 instructions supplied with SNAP-tag plasmids. The labeling of SNAP-tag fusion proteins with SNAP-Surface substrates is described in the instructions supplied with SNAP-Surface substrates. The use of SNAP-Surface Block during the labeling of fusion proteins with SNAP-Surface substrates is described below.
Materials Required but not Supplied
• Cells expressing SNAP-tag fusion proteins
• Tissue culture materials and media
• Transfection reagents
• Fluorescence microscope with suitable filter set
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