标签归档：his

pEF1/myc-His C载体说明书

发表于由

型号 载体名称 出品公司 载体用途 价格
VPI0028 pEF1/myc-His C Invitrogen 哺乳动物表达载体 800

 

产品参数: 

Key Function：Constitutive Expression
Mammalian Selection：Neo，G418
Promoter：EF-1α
Vector Type：pEF
Cloning Method：Restriction Enzyme/MCS 
Protein Tag or Fusion：c-Myc Epitope Tag，His Tag （6x His） 
Constitutive or Inducible System：Constitutive

载体抗性: 

氨苄青霉素（Ampicillin）

载体描述: 

pEF1/myc-His A, B, and C are 6.2 kb vectors derived from pcDNA 3.1/myc-His and designed for overproduction of recombinant proteins in mammalian cell lines. Features of the vectors allow purification and detection of expressed proteins. High-level stable and transient expression can be carried out in most mammalian cells. The vectors contain the following elements:

  1. Human elongation factor 1?-subunit （hEF-1α） promoter for high-level expression across a broad range of species and cell types （Goldman et al., 1996; Mizushima and Nagata, 1990）.
  2. Three reading frames to facilitate in-frame cloning with a C-terminal tag encoding the myc （c-myc） epitope and a polyhistidine metal-binding peptide.
  3. Neomycin resistance gene for selection of stable cell lines.
  4. Episomal replication in cell lines that are latently infected with SV40 or that express the SV40 large T antigen （e.g., COS7）.

The control plasmid, pEF1/myc-His/lacZ, is included for use as a positive control for transfection, expression, and detection in the cell line of choice.

质粒图谱: 

pEF1/myc-His B载体说明书

发表于由

型号 载体名称 出品公司 载体用途 价格
VPI0027 pEF1/myc-His B Invitrogen 哺乳动物表达载体 800

 

产品参数: 

Key Function：Constitutive Expression
Mammalian Selection：Neo，G418
Promoter：EF-1α
Vector Type：pEF
Cloning Method：Restriction Enzyme/MCS 
Protein Tag or Fusion：c-Myc Epitope Tag，His Tag （6x His） 
Constitutive or Inducible System：Constitutive

载体抗性: 

氨苄青霉素（Ampicillin）

载体描述: 

pEF1/myc-His A, B, and C are 6.2 kb vectors derived from pcDNA 3.1/myc-His and designed for overproduction of recombinant proteins in mammalian cell lines. Features of the vectors allow purification and detection of expressed proteins. High-level stable and transient expression can be carried out in most mammalian cells. The vectors contain the following elements:

  1. Human elongation factor 1?-subunit （hEF-1α） promoter for high-level expression across a broad range of species and cell types （Goldman et al., 1996; Mizushima and Nagata, 1990）.
  2. Three reading frames to facilitate in-frame cloning with a C-terminal tag encoding the myc （c-myc） epitope and a polyhistidine metal-binding peptide.
  3. Neomycin resistance gene for selection of stable cell lines.
  4. Episomal replication in cell lines that are latently infected with SV40 or that express the SV40 large T antigen （e.g., COS7）.

The control plasmid, pEF1/myc-His/lacZ, is included for use as a positive control for transfection, expression, and detection in the cell line of choice.

质粒图谱: 

pEF1/myc-His A载体说明书

发表于由

型号 载体名称 出品公司 载体用途 价格
VPI0026 pEF1/myc-His A Invitrogen 哺乳动物表达载体 800

 

产品参数: 

Key Function：Constitutive Expression
Mammalian Selection：Neo，G418
Promoter：EF-1α
Vector Type：pEF
Cloning Method：Restriction Enzyme/MCS 
Protein Tag or Fusion：c-Myc Epitope Tag，His Tag （6x His） 
Constitutive or Inducible System：Constitutive

载体抗性: 

氨苄青霉素（Ampicillin）

载体描述: 

pEF1/myc-His A, B, and C are 6.2 kb vectors derived from pcDNA 3.1/myc-His and designed for overproduction of recombinant proteins in mammalian cell lines. Features of the vectors allow purification and detection of expressed proteins. High-level stable and transient expression can be carried out in most mammalian cells. The vectors contain the following elements:

  1. Human elongation factor 1?-subunit （hEF-1α） promoter for high-level expression across a broad range of species and cell types （Goldman et al., 1996; Mizushima and Nagata, 1990）.
  2. Three reading frames to facilitate in-frame cloning with a C-terminal tag encoding the myc （c-myc） epitope and a polyhistidine metal-binding peptide.
  3. Neomycin resistance gene for selection of stable cell lines.
  4. Episomal replication in cell lines that are latently infected with SV40 or that express the SV40 large T antigen （e.g., COS7）.

The control plasmid, pEF1/myc-His/lacZ, is included for use as a positive control for transfection, expression, and detection in the cell line of choice.

质粒图谱: 

pUB6/V5 His C说明书

发表于由

 

 pUB6/V5 His C

型号 载体名称 出品公司 载体用途
VPI0024 pUB6/V5 His C Invitrogen 哺乳动物表达载体

 

产品参数: 

Key Function：Constitutive Expression
Mammalian Selection：Blasticidin
Promoter：UbC
Vector Type：pUB6
Cloning Method：Restriction Enzyme/MCS 
Protein Tag or Fusion：V5 Epitope Tag，His Tag （6x His） 
Constitutive or Inducible System：Constitutive

载体抗性: 
氨苄青霉素（Ampicillin）
载体描述: 

pUB6/V5-His A, B, and C are 5.5 kb vectors designed for overproduction of recombinant proteins in mammalian cell lines. Features of the vectors allow purification and detection of expressed proteins. High-level stable and transient expression can be carried out in most mammalian cells.

The vectors contain the following elements:

  1. Human ubiquitin C promoter （hUbC） for high-level expression across a broad range of species and cell types （Schorpp et al., 1996; Wulff et al., 1990）.
  2. Three reading frames to facilitate in-frame cloning with a C-terminal peptide encoding the V5 epitope and a polyhistidine （6×His） metal-binding tag.
  3. Blasticidin resistance gene （bsd） for selection of stable cell lines.
  4. Episomal replication in cell lines that are latently infected with SV40 or that express the SV40 large T antigen （e.g. COS-1, COS-7）.

The control plasmid, pUB6/V5-His/lacZ, is included for use as a positive control for transfection, expression, and detection in the cell line of choice.

质粒图谱: 
 

pUB6/V5 His B说明书

发表于由

 pUB6/V5 His B

型号 载体名称 出品公司 载体用途
VPI0023 pUB6/V5 His B Invitrogen 哺乳动物表达载体

 

产品参数: 

Key Function：Constitutive Expression
Mammalian Selection：Blasticidin
Promoter：UbC
Vector Type：pUB6
Cloning Method：Restriction Enzyme/MCS 
Protein Tag or Fusion：V5 Epitope Tag，His Tag （6x His） 
Constitutive or Inducible System：Constitutive

载体抗性: 
氨苄青霉素（Ampicillin）
载体描述: 

pUB6/V5-His A, B, and C are 5.5 kb vectors designed for overproduction of recombinant proteins in mammalian cell lines. Features of the vectors allow purification and detection of expressed proteins. High-level stable and transient expression can be carried out in most mammalian cells.

The vectors contain the following elements:

  1. Human ubiquitin C promoter （hUbC） for high-level expression across a broad range of species and cell types （Schorpp et al., 1996; Wulff et al., 1990）.
  2. Three reading frames to facilitate in-frame cloning with a C-terminal peptide encoding the V5 epitope and a polyhistidine （6×His） metal-binding tag.
  3. Blasticidin resistance gene （bsd） for selection of stable cell lines.
  4. Episomal replication in cell lines that are latently infected with SV40 or that express the SV40 large T antigen （e.g. COS-1, COS-7）.

The control plasmid, pUB6/V5-His/lacZ, is included for use as a positive control for transfection, expression, and detection in the cell line of choice.

质粒图谱: 
 

 

 

 

pUB6/V5 His A说明书

发表于由

 pUB6/V5 His A

型号 载体名称 出品公司 载体用途
VPI0022 pUB6/V5 His A Invitrogen 哺乳动物表达载体
产品参数: 

Key Function：Constitutive Expression
Mammalian Selection：Blasticidin
Promoter：UbC
Vector Type：pUB6
Cloning Method：Restriction Enzyme/MCS 
Protein Tag or Fusion：V5 Epitope Tag，His Tag （6x His） 
Constitutive or Inducible System：Constitutive

载体抗性: 
氨苄青霉素（Ampicillin）
载体描述: 

pUB6/V5-His A, B, and C are 5.5 kb vectors designed for overproduction of recombinant proteins in mammalian cell lines. Features of the vectors allow purification and detection of expressed proteins. High-level stable and transient expression can be carried out in most mammalian cells.

The vectors contain the following elements:

  1. Human ubiquitin C promoter （hUbC） for high-level expression across a broad range of species and cell types （Schorpp et al., 1996; Wulff et al., 1990）.
  2. Three reading frames to facilitate in-frame cloning with a C-terminal peptide encoding the V5 epitope and a polyhistidine （6×His） metal-binding tag.
  3. Blasticidin resistance gene （bsd） for selection of stable cell lines.
  4. Episomal replication in cell lines that are latently infected with SV40 or that express the SV40 large T antigen （e.g. COS-1, COS-7）.

The control plasmid, pUB6/V5-His/lacZ, is included for use as a positive control for transfection, expression, and detection in the cell line of choice.

质粒图谱: 
 

pcDNA3.1（-）/myc-His A&B&C说明书

发表于由

  

pcDNA3.1（-）/myc-His A&B&C

型号 载体名称 出品公司 载体用途
VPI0017 pcDNA3.1（-）/myc-His A Invitrogen 哺乳动物表达载体
VPI0018 pcDNA3.1（-）/myc-His B Invitrogen 哺乳动物表达载体
VPI0019 pcDNA3.1（-）/myc-His C Invitrogen 哺乳动物表达载体

 

产品参数: 

Mammalian Selection：G418，neo
Protein Tag or Fusion：c-Myc Epitope Tag，His Tag （6x His）
Sequencing Primer：T7 Fwd
Sequencing Primer Sequence：5’d[TAATACGACTCACTATAGGG]3′

载体抗性: 
氨苄青霉素（Ampicillin）
质粒图谱: 

 

Invitrogen四环素诱导系统（pCDNA4/HisMax A&B&C）说明书

发表于由

 Invitrogen四环素诱导系统

Vectors：pcDNA4/TO/Myc-His A，pcDNA4/TO/Myc-His B，pcDNA4/TO/Myc-His C，

 

产品参数: 

Mammalian Selection：Zeocin
Sequencing Primer：T7 Fwd
Sequencing Primer Sequence：5’d[TAATACGACTCACTATAGGG]3′ 
Tag：6X His，Xpress

载体抗性: 
氨苄青霉素（Ampicillin）
 
 
质粒图谱: 

 

Invitrogen pGene v5-His 说明书

发表于由

 

GeneSwith 哺乳动物诱导系统

Invitrogen  pGene v5-His 

 

 

产品参数: 

Mammalian Selection：Zeocin
Constitutive/Inducible：Inducible
Expression Level Tightly controlled （activate with mifepristone）
Sequencing Primer pGene Fwd 
Sequencing Primer Sequence：5’d[CTGCTATTCTGCTCAACCT]3′ 
Protein Tags：6X His，V5

载体抗性: 
氨苄青霉素（Ampicillin）
载体描述: 

Switch-On Expression from the Lowest Basal Levels

The GeneSwitch™ System for inducible mammalian expression is ideal for experiments that require the absolute lowest uninduced expression levels. The expression vector pGene/V5-His provides a minimal promoter, GAL4-E1b, consisting of the binding sites for the yeast Gal4 DNA binding protein followed by the TATA sequence from the Adenovirus E1b promoter. Without additional factors, the GAL4-E1b promoter is transcriptionally silent.

To activate transcription from the GAL4-E1b promoter, the GeneSwitch™ regulatory protein is expressed from a minimal TK promoter on the pSwitch vector. The GeneSwitch™ protein has three functional domains:

  1. Gal4 DNA Binding Domain （Gal4-DBD） to bind the regulatory protein to the GAL4-E1b promoter
  2. Truncated human Progesterone Receptor Ligand Binding Domain （hPR-LBD） that undergoes a conformational change when it binds the progesterone antagonist, mifepristone
  3. Transcription activation domain from the NFØB transcription factor p65 （p6D） to activate transcription from the silent GAL4-E1b promoter

In the absence of mifepristone, the conformation of the hPR-LBD region prevents the GeneSwitch™ regulatory protein from activating transcription from the GAL4-E1b promoter. When mifepristone is added and binds the hPR-LBD region, the GeneSwitch™ regulatory protein assumes a conformation that permits it to stimulate transcription from the GAL4-E1b promoter.

Induction of the GeneSwitch™ System leads to activation of the gene of interest on the vector pGene/V5-His. In addition, four Gal4 binding sites upstream of the minimal HSV TK promoter on pSwitch can bind the pSwitch regulatory protein. Therefore, adding mifepristone up-regulates production of the regulatory protein. The increased levels of the GeneSwitch™ regulatory protein result in induction of the gene of interest from pGene/V5-His to levels that can approach those of viral promoters.

The GeneSwitch™ Kit offers exceptionally low uninduced and high induced expression levels in mammalian cells. The system provides the expression vector pGene/V5-His with the minimal, synthetic GAL4-E1b promoter that is transcriptionally silent until activated. The GeneSwitch™ regulatory protein binds the GAL4-E1b promoter to activate transcription upon the addition of mifepristone. In addition, the GeneSwitch™ protein upregulates its own expression, leading to amplified expression of the gene of interest from pGene/V5-His. pGene/V5-His offers several features that facilitate expression analysis and purification of recombinant proteins in mammalian cells:

  1. C-terminal V5 epitope tag for detection with Anti-V5 Antibodies
  2. C-terminal polyhistidine （6xHis） tag for rapid purification with nickel-chelating resin and detection with Anti-His（C-term） Antibodies
  3. The Zeocin resistance gene for effective selection in both mammalian cells and E. coli

 

cube-biotech他的亲和力树脂和磁珠说明书

发表于由

 

cube-biotech他的亲和力树脂和磁珠说明书

多组氨酸标签/Poly-His-标签是应用广泛的蛋白质亲和标签。由于其只有六个氨基酸的小尺寸，它很少干扰蛋白质的功能。此外，它在天然和变性条件下的低免疫原性和多功能性突出了这种蛋白质标签。His-tag 的基本纯化原理是金属离子与组氨酸咪唑环之间的相互作用。

 

His-tag 纯化树脂和磁珠：

选择正确类型的 His-tag 纯化树脂或磁珠基于三个因素：

  1. 配体：这是金属离子在螯合剂复合物中偶联的分子（见图 1）。
    Cube Biotech 提供三种不同的配体。

    • NTA：次氮基三乙酸是常用的配体。它与镍的螯合剂络合物如图 1 所示。

    • IDA：亚氨基二乙酸是第二常用的一种。单击此处了解IDA 和 NTA 之间的区别。

    • INDIGO：这种配体是在 Cube Biotech 开发的。它的优点之一是大大提高了 DTT 和 EDTA 耐受性。单击此处了解有关 INDIGO的更多信息。

  2. 偶联的金属离子：取决于与配体偶联的金属离子，His 标签纯化的亲和力和特异性会发生变化。经验法则：蛋白质产量越高，纯化的特异性越低。图 2 给出了用于 His-tag 纯化的常用金属离子的亲和力/特异性比率的一个概览。

  3. 珠子的大小：使用的珠子直径越大，FPLC 纯化的流速越高。然而，具有较慢的流速通常会增加蛋白质产量。我们提供 30 µm、40 µm、100 µm 和 XL 尺寸（~400 µm）的珠子直径。可根据要求提供更多尺寸。参见图 3 以供参考。

 

 

His-tag 亲和树脂和 MagBeads 的详细信息：

 

图 1：由 NTA 和镍2+组成的螯合剂复合物的示意图。该镍离子与添加到蛋白质中的 His 标签的两个咪唑环结合。
图 2：His 标签蛋白纯化中常用的金属离子概述。铜的亲和力高，而特异性低。钴标志着光谱的另一端。镍是常用的金属离子，在亲和力和特异性之间具有良好的平衡。

PureCube 100 INDIGO 镍琼脂糖

 

 

 

产品信息“PureCube 100 INDIGO Ni-Agarose”
类型： 琼脂糖
珠子大小： 100 µm（琼脂糖）
亲和力： 他的标签
目的： 蛋白质纯化
配体： 靛青
耦合离子： Ni2+


PureCube 100 INDIGO-Ni 琼脂糖树脂是 Cube Biotech 为高质量的 His-tag 蛋白质纯化而开发的产品。其目的是创造一种琼脂糖树脂，能够承受高浓度的螯合剂，如 EDTA 和 DTT，这些螯合剂在哺乳动物细胞培养缓冲液中很常见。得到的 INDIGO 配体具有 20 mM 的 DTT 和 EDTA 公差。这远远优于其他竞争树脂。INDIGO-Ni 也可用作磁珠或预装在墨盒/柱中。