pCLIPf-H2B Control Plasmid

产品编号：N9218S 品牌：New England Biolabs 原厂货号：N9218S
产品分类：克隆 > 克隆载体和克隆试剂盒 > 载体 > DNA亚克隆载体
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包装：

20 μg

运保温度:

-20°C

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描述：

This control plasmid contains the gene encoding the Histone H2B protein cloned upstream of the CLIP_fcoding sequence in pCLIP_f as a fusion to the N- terminus of the CLIP-tag. Histone H2B is a member of the core histones that package DNA in the nucleus. The H2B-CLIP_f fusion protein gives nuclear fluorescence when labeled with CLIP-Cell™ substrates. The full sequence and map for pCLIP_f-H2B can be downloaded.

The CLIP-tag™ protein labeling system enables the specific, covalent attachment of virtually any molecule to a protein of interest. CLIP-tag is a protein tag 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 bond. 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 and complementary labeling of two proteins simultaneously in the same cells.

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.

There are two steps to using this system: sub cloning and expression of the protein of interest as a CLIP_ffusion, and labeling of the fusion with the CLIP-tag substrate of choice. Expression of the CLIP_f-H2B fusion protein is described in this document. The labeling of fusion proteins with CLIP-tag substrates is described in the instructions supplied with CLIP-tag substrates.

注意事项：

Storage: pCLIP_f-H2B 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.