title: Fluorescent labeling of COS-7 expressing SNAP-tag fusion proteins for live cell imaging.
abstract: SNAP-tag and CLIP-tag protein labeling systems enable the specific, covalent attachment of molecules, including fluorescent dyes, to a protein of interest in live cells. These systems offer a broad selection of fluorescent substrates optimized for a range of imaging instrumentation. Once cloned and expressed, the tagged protein can be used with a variety of substrates for numerous downstream applications without having to clone again. There are two steps to using this system: 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. The SNAP-tag is a small protein based on human O(6)-alkylguanine-DNA-alkyltransferase (hAGT), a DNA repair protein. SNAP-tag labels are dyes 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. CLIP-tag is a modified version of SNAP-tag, engineered to react with benzylcytosine rather than benzylguanine derivatives. When used in conjunction with SNAP-tag, CLIP-tag enables the orthogonal and complementary labeling of two proteins simultaneously in the same cells.
authors.affiliation: Division of Chemical Biology, New England Biolabs. provost@neb.com
authors.forename: Christopher R
authors.initials: CR
authors.lastname: Provost
authors.forename: Luo
authors.initials: L
authors.lastname: Sun
mesh_terms: D000818:Animals; D016466:CHO Cells; D019556:COS Cells; D002522:Chlorocebus aethiops; D006224:Cricetinae; D003412:Cricetulus; D005456:Fluorescent Dyes / Q000737:chemistry*; D007089:Image Enhancement / Q000379:methods*; D008856:Microscopy, Fluorescence / Q000379:methods; D019853:O(6)-Methylguanine-DNA Methyltransferase / Q000096:biosynthesis / Q000737:chemistry / Q000235:genetics; D011993:Recombinant Fusion Proteins / Q000096:biosynthesis / Q000737:chemistry* / Q000235:genetics; D014162:Transfection
chemical_list: D005456:Fluorescent Dyes; D011993:Recombinant Fusion Proteins; D019853:O(6)-Methylguanine-DNA Methyltransferase