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          生物通商城首页 > 试剂 > 分子生物学 > 报告系统/蛋白标记定位 > CLIP-Cell

          CLIP-Cell 启动试剂盒

          CLIP-Cell Starter Kit, 1 set

          品牌:NEB 货号:E9200S 立即咨询 QQ 咨询留言

          生物通价:

          ¥2294.00目录价:¥2699.00

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          ¥2294.00
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          • 基因有限公司 发货并提供售后服务

          Shelf Life(month):24
          产地:USA
          存储条件:-20
          蛋白质标记系统启动试剂盒提供了标记CLIP-tag融合蛋白的所有必需组份。能在活细胞、固定细胞及体外将红色或绿色荧光基团共价连接到目标蛋白上。每种启动试剂盒包括一个编码所选 tag 的质粒和两种细胞通透性的荧光标记物。试剂盒中还提供一种阳性对照质粒,其编码有亚细胞定位明确的标签蛋白(如:膜蛋白、核蛋白等)。如果必要,试剂盒还会提供一个与目的标签相互作用的、非荧光的阴性对照阻断剂。The SNAP-tag® and CLIP-tag™ are novel tools for the specific, covalent attachment of virtually any molecule to a protein of interest, providing simplicity and extraordinary versatility to the imaging of proteins in live and fixed cells, and to the study of proteins in vitro. The creation of a single gene construct yields a tagged fusion protein capable of forming a covalent linkage to a variety of functional groups, including fluorophores, biotin, or beads. This system provides a powerful and unique tool to study the role of proteins in a variety of highly dynamic processes, including protein trafficking, turnover and complex formation. The CLIP-tag is a 20 kDa mutant of the human DNA repair protein O6-alkylguanine- DNA alkyltransferase (hAGT) that reacts specifically and rapidly with benzylcytosine (BC) derivatives, leading to covalent labeling of the CLIP-tag with a synthetic probe (Figure 1).

          The CLIP-tag has a number of features that make it ideal for a variety of protein labeling applications. The rate of the reaction of the CLIP-tag with these derivatives is largely independent of the nature of the synthetic probe attached to BC, permitting the labeling of CLIP fusion proteins with a wide variety of functional groups. Many of these CLIP-tag substrates are cell-permeable, allowing live-cell imaging of protein expression and localization (Figure 2). The ability to turn on the signal at will, together with the availability of a cell-permeable nonfluorescent blocking agent (CLIP-Cell™ Block), allows time-resolved pulse-chase analysis of protein trafficking. Finally, the availability of orthogonal protein labeling systems from NEB permits simultaneous labeling of multiple proteins in a single cell (SNAP-tag, another hAGT variant that reacts exclusively with O6-benzylguanine substrates, and the ACP/MCP tags, small protein tags which can be enzymatically labeled on the cell surface with Coenzyme A derivatives).

          The CLIP-Cell Starter Kit contains a mammalian expression plasmid (pCLIPf) encoding the CLIP-tag flanked by restriction sites for cloning a gene of interest, and two cell-permeable fluorescent CLIP-tag substrates. A positive control plasmid (pCLIPf-H2B), encoding a CLIP-tagged protein (histone H2B) with a well-characterized nuclear localization, is also included. Lastly, a negative control “blocking agent” (CLIP-Cell Block) is included that interacts with the CLIP-tag, but is not fluorescent. There are two steps to using this system: subcloning and expression of the protein of interest as a CLIPf fusion, and labeling of the fusion with the CLIP-tag substrate of choice.
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