CLIP-Biotin is a cell-permeable substrate (BC-Biotin) based on biotin with an amidocaproyl linker. It is suitable for applications such as biotinylation of CLIP-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 CLIP-Biotin substrate, sufficient to make 10 ml of a 5 μM solution for the labeling of CLIP-tag fusion proteins in cells.
CLIP-tag is a novel tool for protein research, allowing the specific, covalent attachment of virtually any molecule to a protein of interest. CLIP-tag is a small protein based on mammalian O6-alkylguanine-DNA-alkyltransferase (AGT). CLIP-tag substrates are derivatives of benzylcytosine (BC). In the labeling reaction, the substituted benzyl group of the substrate is covalently attached to the reactive cysteine of CLIP-tag forming a stable thioether link. Although CLIP-tag is based on the same protein as SNAP-tag®, the benzylcytosine substrates form a separate class of substrates, different from the benzylguanine substrates recognized by SNAP-tag. CLIP-tag and SNAP-tag can be used for orthogonal simultaneous labeling.
There are two steps to using this system: subcloning and expression of the protein of interest as a CLIP-tag fusion, and labeling of the fusion with the CLIP-tag substrate of choice. Expression of CLIP-tag fusion proteins is described in the documentation supplied with CLIP-tag plasmids. The labeling of fusion proteins with CLIP-Biotin is described below.
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