pCLIPf Vector

产品编号：N9215S 品牌：New England Biolabs 原厂货号：N9215S
产品分类：克隆 > 克隆载体和克隆试剂盒 > 载体 > 哺乳动物表达载体
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-20°C

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描述：

pCLIP_f Vector is a mammalian expression plasmid intended for the cloning and stable or transient expression of CLIP-tag® protein fusions in mammalian cells. This plasmid encodes CLIP_f, a CLIP-tag protein, which is expressed under control of the CMV promoter. The expression vector has an IRES (internal ribosome entry site) and a neomycin resistance gene downstream of the CLIP_f for the efficient selection of stable transfectants. pCLIP_f Vector contains two multiple cloning sites to allow cloning of the fusion partner as a fusion to the N- or C-terminus of the CLIP_f.

The CLIP-tag is a novel tool for protein research, allowing the specific, covalent attachment of virtually any molecule to a protein of interest. The CLIP-tag is a small protein based on human O⁶-alkylguanine-DNA-alkyltransferase (hAGT). 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.

pCLIP_f contains an improved version of CLIP-tag, termed CLIP_f. CLIP_f displays faster kinetics in in vitrolabeling and fast, specific and efficient labeling in live and fixed cell applications, thereby rendering it a desired research tool for analysis of protein dynamics.

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: sub cloning and expression of the protein of interest as a CLIP_f fusion, and labeling of the fusion with the CLIP-tag substrate of choice. Cloning and expression of CLIP_f fusion proteins are described in this document. The labeling of the fusion proteins with CLIP-tag substrates is described in the instructions supplied with the CLIP-tag substrates.

注意事项：

Storage: pCLIP_f Vector is supplied in TE buffer (10 mM Tris-HCl, pH 8.0, 1 mM EDTA) at a concentration of 0.5 µg/µl. Plasmid solutions can be stored at 4°C for up to one week. For long-term storage -20°C is recommended.