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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 > SNAP-Cell

          SNAP-Surface 启动试剂盒

          SNAP-Surface Starter Kit

          品牌:NEB 货号:E9120S 立即咨询 QQ 咨询留言

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          ¥2294.00目录价:¥2699.00

          生物通价:

          ¥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 SNAP-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编码SNAP-tag的质粒、两种光稳定的非细胞通透性荧光底物SNAP-Surface 488(绿色)和SNAP-Surface 549(红色),阻断剂和对照,可在活细胞表面或溶液/体外对目标蛋白进行荧光示踪。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-Surface Starter Kit contains a mammalian expression plasmid (pSNAPf) encoding the SNAP-tag flanked by restriction sites for cloning a gene of interest, and two non-cell-permeable fluorescent SNAP-tag substrates. A positive control plasmid (pSNAPf-ADRβ2), encoding a SNAP-tagged protein (beta-2 adrenergic receptor) with a well-characterized cell surface localization, is also included. Lastly, a negative control “blocking agent” (SNAP-Surface Block) is included that interacts with the SNAP-tag, but is not fluorescent. There are two steps to using this system: subcloning and expression of the protein of interest as a SNAPf fusion, and labeling of the fusion with the SNAP-tag substrate of choice.

          The SNAP-tag® is a novel tool 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 SNAP-tag is a 20 kDa mutant of the human DNA repair protein O6-alkylguanine- DNA alkyltransferase (hAGT) that reacts specifically and rapidly with benzylguanine (BG) and benzylchloropyrimidine (CP) derivatives, leading to covalent labeling of the SNAP-tag with a synthetic probe (Figure 1). The SNAP-tag has a number of features that make it ideal for a variety of protein labeling applications. The rate of the reaction of the SNAP-tag with these derivatives is largely independent of the nature of the synthetic probe attached to BG, permitting the labeling of SNAP fusion proteins with a wide variety of functional groups. Many of these SNAP-tag substrates are cell-impermeable, allowing live-cell imaging of protein expression and localization on the cell surface (Figure 2). The ability to turn on the signal at will, together with the availability of a cell-impermeable nonfluorescent blocking agent (SNAP-Surface® Block), allows time-resolved pulse-chase analysis of protein trafficking to the cell surface, as well as subsequent internalization. Finally, the availability of orthogonal protein labeling systems from NEB permits simultaneous labeling of multiple proteins in a single cell (CLIP-tag™, a SNAP-tag variant that reacts exclusively with O2-benzylcytosine substrates, and the ACP/MCP tags, small protein tags which can be enzymatically labeled on the cell surface with Coenzyme A derivatives).
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