SNAP-Cell® 430 is a blue fluorescent substrate that can be used to label SNAP-tag®fusion proteins inside living cells, on cell surfaces, orin vitro. This cell-permeable substrate (BG-430) is based on diethylaminocoumarin and is suitable for appropriate blue lasers and filter sets. It has an excitation maximum at 421 nm and emission maxima at 444 and 484 nm. This package includes 50 nmol of SNAP-Cell 430 substrate, sufficient to make 10 ml of a 5 µM SNAP-tag fusion protein labeling solution.
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 documentation supplied with SNAP-tag plasmids. The labeling of the fusion proteins with the SNAP-tag substrate is described below.