SNAP-Cell® Oregon Green® is a photostable green 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-Oregon Green) is based on the Invitrogen dye, Oregon Green® and is suitable for standard fluorescein filter sets. It has an excitation maximum at 490 nm and an emission maximum at 514 nm. Conjugates of Oregon Green are more photostable than those of fluorescein, and their fluorescence properties are essentially pH insensitive in the physiological pH range. This package contains 50 nmol of SNAP-Cell Oregon Green substrate, sufficient to make 10 ml of a 5 uM 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: 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 the fusion proteins with the SNAP-tag substrate is described below.
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