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SNAP-Cell 阻斷劑:SNAP-Cell Block, 100 nmol Kit

價格:¥

貨号:S9106S
品牌:NEB

經銷:基因生物技術國際貿易(上海)有限公司

阻斷劑是非熒光底物,在細胞内(SNAP-Cell Block)阻斷 SNAP-tag 的反應。在 SNAP-tag 融合蛋白的活細胞标記實驗中,可以用阻斷劑制備無熒光對照。SNAP-Cell 阻斷劑可順利透過細胞膜,一旦進入細胞,即與 SNAP-tag 反應,使之在後續的标記反應中不可逆地失活。SNAP-Cell® Block (bromothenylpteridine, BTP) is a non-fluorescent compound that blocks the reactivity of the SNAP-tag® in solution or in living cells. It can be used to generate inactive controls in live cell labeling experiments performed with SNAP-tag fusion proteins. SNAP-Cell Block is highly membrane permeable and once in the cell reacts with the SNAP-tag, irreversibly inactivating it for subsequent labeling steps.

The SNAP-tag protein labeling system enables the specific, covalent attachment of virtually any molecule to a protein of interest. The SNAP-tag is a small protein based on mammalian O6-alkylguanine-DNA alkyltransferase (AGT). SNAP-tag substrates are derivatives of benzylpurines and benzylchloropyrimidines. In the labeling reaction, the substituted benzyl group of the substrate is covalently attached to the SNAP-tag.

There are two steps to using this system: sub-cloning and expression of the protein of interest as a SNAP-tag fusion, and labeling of the fusion with the SNAP-tag substrate of choice. Expression of SNAP-tag fusion proteins is described in the instructions supplied with SNAP-tag plasmids. The labeling of SNAP-tag fusion proteins with SNAP-Cell substrates is described in the instructions supplied with SNAP-Cell substrates. The use of SNAP-Cell Block during the labeling of fusion proteins with SNAP-Cell substrates is described below.

SNAP-Cell 啟動試劑盒:SNAP-Cell Starter Kit, 1 set Kit

價格:¥

貨号:E9100S
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 SNAP-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 SNAP-Cell Starter Kit contains a mammalian expression plasmid (pSNAPf) encoding the SNAP-tag flanked by restriction sites for cloning a gene of interest, and two cell-permeable fluorescent SNAP-tag substrates. A positive control plasmid (pSNAPf-Cox8A), encoding a SNAP-tagged protein (cytochrome c oxidase) with a well-characterized mitochondrial localization, is also included. Lastly, a negative control “blocking agent” (SNAP-Cell 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-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 (SNAP-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 (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).

SNAP-Cell 505-Star, 50 nmol Kit

價格:¥

貨号:S9103S
品牌:NEB

經銷:基因生物技術國際貿易(上海)有限公司

細胞通透性底物(SNAP-Cell)不僅能标記細胞内部也能标記細胞表面。激發光504nm,發射光532 nm。SNAP-Cell® 505-Star is a photostable green fluorescent substrate that can be used to label SNAP-tag® fusion proteins inside living cells, on cell surfaces, or in vitro. This cell-permeable substrate (CP-6-505) is based on the single isomer 6-carboxyrhodamine 110 and is suitable for standard fluorescein filter sets. It has an excitation maximum at 504 nm and an emission maximum at 532 nm. This package contains 50 nmol of SNAP-Cell 505-Star substrate, sufficient to make 10 ml of a 5 µM SNAP-tag fusion protein labeling solution.

The SNAP-tag protein labeling system enables the specific, covalent attachment of virtually any molecule to a protein of interest. The SNAP-tag is a protein based on human O6-alkylguanine-DNA alkyltransferase (hAGT). SNAP-tag substrates are fluorophores, biotin or beads conjugated to guanine or chloropyrimidine leaving groups via a benzyl linker. In the labeling reaction, the substituted benzyl group of the substrate is covalently attached to the SNAP-tag.

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

SNAP-Cell 430, 50 nmol Kit

價格:¥

貨号:S9109S
品牌:NEB

經銷:基因生物技術國際貿易(上海)有限公司

細胞通透性底物(SNAP-Cell)不僅能标記細胞内部也能标記細胞表面。激發光421nm,發射光444和484 nm。SNAP-Cell® 430 is a blue fluorescent substrate that can be used to label SNAP-tag® fusion proteins inside living cells, on cell surfaces, or in vitro. This cell-permeable substrate (BG-430) is based on diethylaminocoumarin and is suitable for appropriate blue lasers and filter sets. It has an excitation maximum at 421 nm and emission maxima at 444 and 484 nm. This package includes 50 nmol of SNAP-Cell 430 substrate, sufficient to make 10 ml of a 5 µM SNAP-tag fusion protein labeling solution.

The SNAP-tag protein labeling system enables the specific, covalent attachment of virtually any molecule to a protein of interest. The SNAP-tag is a small protein based on human O6-alkylguanine-DNA alkyltransferase (AGT). SNAP-tag substrates are derivatives of benzylpurines and benzylchloropyrimidines. In the labeling reaction, the substituted benzyl group of the substrate is covalently attached to the SNAP-tag.

There are two steps to using this system: sub-cloning and expression of the protein of interest as a SNAP-tag fusion, and labeling of the fusion with the SNAP-tag substrate of choice. Expression of SNAP-tag fusion proteins is described in the documentation supplied with SNAP-tag plasmids. The labeling of the fusion proteins with the SNAP-tag substrate is described below.

SNAP-Cell TMR-Star, 30 nmol Kit

價格:¥

貨号:S9105S
品牌:NEB

經銷:基因生物技術國際貿易(上海)有限公司

細胞通透性底物(SNAP-Cell)不僅能标記細胞内部也能标記細胞表面。激發光554nm,發射光580nm。SNAP-Cell® TMR-Star is a red fluorescent substrate that can be used to label SNAP-tag® fusion proteins inside living cells, on cell surfaces, or in vitro. This cell-permeable substrate (CP-6-TMR) is based on the single isomer 6-carboxytetramethylrhodamine and is suitable for standard rhodamine filter sets. It has an excitation maximum at 554 nm and an emission maximum at 580 nm. This package contains 30 nmol of SNAP-Cell TMR-Star substrate, sufficient to make 10 ml of a 3 µM SNAP-tag fusion protein labeling solution.

The SNAP-tag protein labeling system enables the specific, covalent attachment of virtually any molecule to a protein of interest. The SNAP-tag is a small protein based on mammalian O6-alkylguanine-DNA alkyltransferase (AGT). SNAP-tag substrates are derivatives of benzylpurines and benzylchloropyrimidines. In the labeling reaction, the substituted benzyl group of the substrate is covalently attached to the SNAP-tag.

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

SNAP-Cell 647-SiR, 30 nmol Kit

價格:¥

貨号:S9102S
品牌:NEB

經銷:基因生物技術國際貿易(上海)有限公司

細胞通透性底物(SNAP-Cell)不僅能标記細胞内部也能标記細胞表面。激發光 645nm,發射光661nm。

SNAP-Cell 647-SiR is a far-red fluorescent substrate that can be used to label SNAP-tag® fusion proteins inside living cells, on cell surfaces, or in vitro. This cell-permeable substrate (also termed SiR-SNAP)(1) is based on 6-carboxy-tetramethylsiliconrhodamine and is suitable for standard Cy5 filter sets. It has an excitation maximum at 645 nm and an emission maximum at 661 nm. This package contains 30 nmol of SNAP-Cell 647-SiR substrate, sufficient to make 10 ml of a 3 μM SNAP-tag fusion protein labeling solution.

The SNAP-tag protein labeling system enables the specific, covalent attachment of virtually any molecule to a protein of interest. The SNAP-tag is based on human O6-alkylguanine-DNA-alkyltransferase (hAGT). SNAP-tag substrates are fluorophores, biotin or beads conjugated to guanine or chloropyrimidine leaving groups via a benzyl linker. In the labeling reaction, the substituted benzyl group of the substrate is covalently attached to the SNAP-tag.

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

SNAP-Cell Oregon Green®:SNAP-Cell Oregon Green, 50 nmol Kit

價格:¥

貨号:S9104S
品牌:NEB

經銷:基因生物技術國際貿易(上海)有限公司

細胞通透性底物(SNAP-Cell)不僅能标記細胞内部也能标記細胞表面。激發光490 nm,發射光514 nm。SNAP-Cell Oregon Green is a photostable green fluorescent substrate that can be used to label SNAP-tag® fusion proteins inside living cells. This cell-permeable substrate (BG-Oregon Green) is based on the Invitrogen dye, Oregon Green and is suitable for standard fluorescein filter sets. It has an excitation maximum at 490 nm and an emission maximum at 514 nm. Conjugates of Oregon Green are more photostable than those of fluorescein, and their fluorescence properties are essentially pH insensitive in the physiological pH range. This package contains 50 nmol of SNAP-Cell Oregon Green substrate, sufficient to make 10 ml of a 5 µM SNAP-tag fusion protein labeling solution.

The SNAP-tag is a novel tool for protein research, allowing the specific, covalent attachment of virtually any molecule to a protein of interest. The SNAP-tag is based on mammalian O6-alkylguanine-DNA-alkyltransferase (AGT). SNAP-tag substrates are derivatives of benzylguanines and benzylchloropyrimidines. In the labeling reaction, the dye-substituted benzyl group of the substrate becomes covalently attached to the SNAP-tag.

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

BG-NH2, 2 mg Kit

價格:¥

貨号:S9148S
品牌:NEB

經銷:基因生物技術國際貿易(上海)有限公司

SNAP-tag底物,苄基鳥嘌呤(benz ylguanine,BG),用于連接NHS酯和其他含活化羧基的酯。可用于将新的分子或配基連至蛋白質;制作用于标記蛋白的自定義底物;制作用于蛋白質固定的載體表面。如耦聯至 Biacore® 芯片或微陣列表面,以固定特定的蛋白質;還可耦聯至多肽、蛋白質或DNA寡聚物。耦聯至新的熒光基團或親和試劑,可以标記特定的蛋白質。标記反應溫和、精确且作用于多種底物:在生物學條件下,标記物可以在特定位置形成共價鍵。

BG-PEG-NH2, 2 mg Kit

價格:¥

貨号:S9150S
品牌:NEB

經銷:基因生物技術國際貿易(上海)有限公司

SNAP-tag底物,苄基鳥嘌呤(benz ylguanine,BG),PEG-linker極大提高了柔韌性,特别适合将蛋白固定在固體表面。可用于将新的分子或配基連至蛋白質;制作用于标記蛋白的自定義底物;制作用于蛋白質固定的載體表面。如耦聯至 Biacore® 芯片或微陣列表面,以固定特定的蛋白質;還可耦聯至多肽、蛋白質或DNA寡聚物。耦聯至新的熒光基團或親和試劑,可以标記特定的蛋白質。标記反應溫和、精确且作用于多種底物:在生物學條件下,标記物可以在特定位置形成共價鍵。

BG-GLA-NHS, 2 mg Kit

價格:¥

貨号:S9151S
品牌:NEB

經銷:基因生物技術國際貿易(上海)有限公司

SNAP-tag底物,苄基鳥嘌呤(benz ylguanine,BG)活化為NHS酯,與伯胺反應。可用于将新的分子或配基連至蛋白質;制作用于标記蛋白的自定義底物;制作用于蛋白質固定的載體表面。如耦聯至 Biacore® 芯片或微陣列表面,以固定特定的蛋白質;還可耦聯至多肽、蛋白質或DNA寡聚物。耦聯至新的熒光基團或親和試劑,可以标記特定的蛋白質。标記反應溫和、精确且作用于多種底物:在生物學條件下,标記物可以在特定位置形成共價鍵。

BG-Maleimide, 2 mg Kit

價格:¥

貨号:S9153S
品牌:NEB

經銷:基因生物技術國際貿易(上海)有限公司

SNAP-tag底物,苄基鳥嘌呤(benz ylguanine,BG)活化為馬來酰亞胺,與硫醇類物質反應。可用于将新的分子或配基連至蛋白質;制作用于标記蛋白的自定義底物;制作用于蛋白質固定的載體表面。如耦聯至 Biacore® 芯片或微陣列表面,以固定特定的蛋白質;還可耦聯至多肽、蛋白質或DNA寡聚物。耦聯至新的熒光基團或親和試劑,可以标記特定的蛋白質。标記反應溫和、精确且作用于多種底物:在生物學條件下,标記物可以在特定位置形成共價鍵。

SNAP-Surface 啟動試劑盒:SNAP-Surface Starter Kit 1 set

價格:¥

貨号:E9120S
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 SNAP-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編碼SNAP-tag的質粒、兩種光穩定的非細胞通透性熒光底物SNAP-Surface 488(綠色)和SNAP-Surface 549(紅色),阻斷劑和對照,可在活細胞表面或溶液/體外對目标蛋白進行熒光示蹤。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 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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