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生物通商城首页 -> 试剂 -> 细胞生物学 -> 蛋白质标记定位 -> CLIP-tag系统

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SNAP-tag 和 CLIP-tag 蛋白标记系统能将目标蛋白以共价结合的形式,特异性地标记上任何分子。使用该系统时应包括两个步骤:亚克隆目标蛋白质,使其与 SNAP-tag 形成融合表达蛋白;选择理想底物标记SNAP-tag。SNAP-tag 是一种分子量较小的多肽,来自 DNA 修复蛋白人烷基鸟嘌呤-DNA 烷基转移酶(O6-alkylguanine-DNA-alkyltransferase,hAGT),SNAP-tag的底物可以是通过苄基与鸟嘌呤或氯嘧啶基团偶联的荧光基团、生物素或微珠。在标记反应中,底物的苄基部份被替换,与 SNAP-tag 形成共价结合。CLIP-tag 是 SNAP-tag 的改良版本,与苄基胞嘧啶反应,而不是与苄基鸟嘌呤的衍生物反应。CLIP-tag与 SNAP-tag 共同使用时,能在同一细胞中同时标记两种蛋白。

 

CLIP-Cell™:CLIP-Cell 标记物是细胞通透性底物,适用于在活细胞内、固定细胞内、活细胞表面、固细胞表面或体外标记携带有 CLIP-tag 的融合蛋白。CLIPtag由 SNAP-tag 衍生而来,与不同底物特异性结合,可以同时以不同荧光底物标记两种表达蛋白。该类产品的阻断剂是具有细胞通透性的非荧光标记物。

 

CLIP-Surface™:CLIP-Surface 标记物是细胞非通透性底物,适用于在活细胞表面、固定细胞表面或体外标记携带有 CLIP-tag 的融合蛋白。CLIP-tag 由SNAP-tag 衍生而来,与不同底物特异性结合,可以同时以不同荧光底物标记两种表达蛋白。这类标记物包括了常用可见光谱区域(如 488、547 和647 nm)的探针。

 

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产品名/货号/品牌 简介 包装  
CLIP-Cell Block, 100 nmol 100 nmol

价格:¥

货号:S9220S
品牌:NEB

经销:基因有限公司

非荧光底物,可顺利透过细胞膜,在细胞内或活细胞表面阻断SNAP-tag的反应,使之在后续的标记反应中不可逆地失活。在Clip-tag融合蛋白的活细胞标记实验中,可以用阻断剂制备无荧光对照

pCLIPf Vector, 20 μg 20 ug

价格:¥

货号:N9215S
品牌:NEB

经销:基因有限公司

CLIP-Cell Starter Kit, 1 Kit 1 Kit

价格:¥

货号:E9200S
品牌:NEB

经销:基因有限公司

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.

CLIP-Surface Starter Kit, 1 Kit 1 Kit

价格:¥

货号:E9230S
品牌:NEB

经销:基因有限公司

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 的质粒和细胞非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 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-Biotin , 50 nmol 50 nmol

价格:¥

货号:S9221S
品牌:NEB

经销:基因有限公司

CLIP-Cell 505 , 50 nmol 50 nmol

价格:¥

货号:S9217S
品牌:NEB

经销:基因有限公司

细胞通透性底物(CLIP-Cell)激发波长504nm, 发射波长532nm。CLIP-Cell™ 505 is a green fluorescent substrate that can be used to label CLIP-tag™ fusion proteins inside living cells, on cell surfaces or in vitro. This cell-permeable substrate (BC-505) is based on the Dyomics dye DY-505 and is suitable for standard fluorescein filter sets. It has an excitation maximum at 504 nm and emission maximum at 532 nm. This package includes 50 nmol of CLIP-Cell 505 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-DNAalkyltransferase (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.

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

CLIP-Cell TMR-Star , 30 nmol 30 nmol

价格:¥

货号:S9219S
品牌:NEB

经销:基因有限公司

细胞通透性底物(CLIP-Cell)激发波长554nm, 发射波长580nm。CLIP-Cell™ TMR-Star is a photostable red fluorescent substrate that can be used to label CLIP-tag™ fusion proteins inside living cells, on cell surfaces or in vitro. This cell-permeable substrate (BC-TMR) is based on tetramethylrhodamine and is suitable for standard rhodamine filter sets. It has an excitation maximum at 554 nm and emission maximum at 580 nm. This package includes 30 nmol of CLIP-Cell TMR-Star, sufficient to make 10 ml of a 3 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-DNAalkyltransferase (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.

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 with the CLIP-tag substrate is described in this document.

CLIP-Surface 488 , 50 nmol 50 nmol

价格:¥

货号:S9232S
品牌:NEB

经销:基因有限公司

细胞非通透性底物(CLIP-Surface)激发波长506nm, 发射波长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 50 nmol

价格:¥

货号:S9233S
品牌:NEB

经销:基因有限公司

细胞非通透性底物(CLIP-Surface)激发波长554nm, 发射波长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 50 nmol

价格:¥

货号:S9234S
品牌:NEB

经销:基因有限公司

细胞非通透性底物(CLIP-Surface)激发波长660nm, 发射波长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.

BC-NH2 2 mg

价格:¥

货号:S9236S
品牌:NEB

经销:基因有限公司

CLIP-tag底物,苄基胞嘧啶(benzylcytosine,BC),用于连接NHS酯和其他含活化羧基的酯。可用于将新的分子或配基连至蛋白质;制作用于标记蛋白的自定义底物;制作用于蛋白质固定的载体表面。如耦联至 Biacore® 芯片或微阵列表面,以固定特定的蛋白质;还可耦联至多肽、蛋白质或DNA寡聚物。耦联至新的荧光基团或亲和试剂,可以标记特定的蛋白质。标记反应温和、精确且作用于多种底物:在生物学条件下,标记物可以在特定位置形成共价键。

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