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CLIP-Surface 啟動試劑盒:CLIP-Surface Starter Kit, 1 set Kit

價格:¥

貨号:E9230S
品牌:NEB

經銷:基因生物技術國際貿易(上海)有限公司

蛋白質标記系統啟動試劑盒提供了标記CLIP-tag融合蛋白的所有必需組份。能在活細胞、固定細胞及體外将紅色或綠色熒光基團共價連接到目标蛋白上。每種啟動試劑盒包括一個編碼所選 tag 的質粒和細胞非通透性的熒光标記物。試劑盒中還提供一種陽性對照質粒,其編碼有亞細胞定位明确的标簽蛋白(如:膜蛋白、核蛋白等)。如果必要,試劑盒還會提供一個與目的标簽相互作用的、非熒光的陰性對照阻斷劑。The CLIP-Surface Starter Kit contains a mammalian expression plasmid (pCLIPf) encoding the CLIP-tag flanked by restriction sites for cloning a gene of interest, and two non-cell-permeable fluorescent CLIP-tag substrates. A positive control plasmid (pCLIPf-NK1R), encoding a CLIP-tagged protein (neurokinin-1 receptor) with a well-characterized cell surface 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.

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 non-cell-permeable, 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 nonfluorescent blocking agent (CLIP-Cell™ 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 (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).

CLIP-Surface 488, 50 nmol Kit

價格:¥

貨号:S9232S
品牌:NEB

經銷:基因生物技術國際貿易(上海)有限公司

細胞非通透性底物(CLIP-Surface)激發波長506nm, 發射波長526nm。CLIP-Surface™ 488 is a photostable green fluorescent substrate that can be used to label CLIP-tag™ fusion proteins on the surface of living cells or in vitro. This cell impermeable substrate (BC-488) is based on ATTO-TEC dye ATTO 488 and is suitable for standard fluorescein filter sets. It has an excitation maximum at 506 nm and emission maximum at 526 nm. This package includes 50 nmol of CLIP-Surface 488 substrate, sufficient to make 10 ml of a 5 µM CLIP-tag™ fusion protein labeling solution.

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 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.

There are two steps to using this system: subcloning and expression of the protein of interest as a CLIP-tag fusion, and labeling of the fusion with the CLIP-tag substrate of choice. Expression of CLIP-tag fusion proteins is described in the documentation supplied with CLIP-tag plasmids. The labeling of the fusion proteins on the cell surface with the CLIP-tag substrate is described in this document.

CLIP-Surface 547, 50 nmol Kit

價格:¥

貨号:S9233S
品牌:NEB

經銷:基因生物技術國際貿易(上海)有限公司

細胞非通透性底物(CLIP-Surface)激發波長554nm, 發射波長568nm。CLIP-Surface™ 547 is a photostable red fluorescent substrate that can be used to label CLIP-tag™ fusion proteins on the surface of living cells or in vitro. This cell impermeable substrate (BC-547) is based on the Dyomics dye DY-547 and is suitable for use with standard TAMRA or Cy3 filter sets. It has an excitation maximum at 554 nm and emission maximum at 568 nm. This package includes 50 nmol of CLIP-Surface 547 substrate, sufficient to make 10 ml of a 5 µM CLIP-tag fusion protein labeling solution.

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 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.

There are two steps to using this system: subcloning and expression of the protein of interest as a CLIP-tag fusion, and labeling of the fusion with the CLIP-tag substrate of choice. Expression of CLIP-tag fusion proteins is described in the documentation supplied with CLIP-tag plasmids. The labeling of the fusion proteins on the cell surface with the CLIP-tag substrate is described in this document.

CLIP-Surface 647, 50 nmol Kit

價格:¥

貨号:S9234S
品牌:NEB

經銷:基因生物技術國際貿易(上海)有限公司

細胞非通透性底物(CLIP-Surface)激發波長660nm, 發射波長673nm。CLIP-Surface™ 647 is a photostable red fluorescent substrate that can be used to label CLIP-tag™ fusion proteins on the surface of living cells, or in vitro. This cell impermeable substrate (BC-647) is based on the Dyomics dye DY-647 and is suitable for 635 nm and 650 nm diode laser excitation or use with Cy5 filter sets. It has an excitation maximum at 660 nm and emission maximum at 673 nm. This package includes 50 nmol of CLIP-Surface 647 substrate, sufficient to make 10 ml of a 5 µM CLIP-tag fusion protein labeling solution.

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 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.

There are two steps to using this system: subcloning and expression of the protein of interest as a CLIP-tag fusion, and labeling of the fusion with the CLIP-tag substrate of choice. Expression of CLIP-tag fusion proteins is described in the documentation supplied with CLIP-tag plasmids. The labeling of the fusion proteins on the cell surface with the CLIP-tag substrate is described in this document.

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