pCLIPf-NK1R Control Plasmid

• 产品编号：N9216S      品牌：New England Biolabs       原厂货号：N9216S
• 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
• 应用分类：

描述：

This control plasmid contains the gene encoding the seven-pass transmembrane protein Neurokinin-1 receptor (NK1R), a member of the G-protein coupled receptor family. The NK1R was cloned downstream of the CLIP_f coding sequence in pCLIP_f, as a fusion to the C- terminus of the CLIP_f.

An import signal sequence for import into the endoplasmic reticulum (ER) was cloned upstream of the CLIP_f. This import signal sequence is based on the serotonin receptor 5HT3A. The CLIP_f-neurokinin-1 receptor is inserted in the plasma membrane with the CLIP_f exposed to the extracellular side of the membrane. The CLIP_f-NK1R fusion protein gives plasma membrane labeling when labeled with cell permeable CLIP-Cell™ substrates and non-cell permeable CLIP-Surface substrates.

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_f fusion, and labeling of the fusion with the CLIP-tag substrate of choice. Expression of CLIP_f-NK1R fusion protein is described in this document. The labeling of the fusion proteins with CLIP-tag substrates is described in the instructions supplied with CLIP-tag substrates.

原厂资料：

注意事项：

Storage: pCLIP_f-NK1R 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.