pCMV-3Tag Vectors

pCMV-3Tag Vectors

  • ウエスタン・ブロッティングなどの免疫学的測定法でより高いシグナル強度
  • タンパク質の機能を損なわない短いタグ
  • N-末端、C-末端融合ベクターともに3つのリーディング・フレームを用意
 
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3コピーのFLAG®あるいはc-Mycエピトープ・タグ

エピトープ・タグは、それぞれの目的タンパク質に対して特異的な抗体を作製することなく、それらの検出、追跡、同定を行うことができ、タンパク質の機能解析を行う際に非常に有効なツールです。アジレント(旧ストラタジーン)のpCMV-3Tag Vectorシリーズは、目的のタンパク質のN-末端もしくはC-末端へFLAG®あるいはc-Mycのいずれかのタグを3コピー付けられる、哺乳類細胞での発現ベクター・システムです。FLAG®とc-Mycはそれぞれ8アミノ酸(DYKDDDDK)と 10アミノ酸(EQKLISEEDL)と小さいため、融合させる目的のタンパク質の機能を損なうことがほとんどありません。また、免疫反応性が非常に高く、市販されている抗体によって簡単に検出することができます。また、タグを3コピー付加することにより、ウェスタン・ブロッティング、in situ染色、免疫沈降法などの免疫学的測定法でのシグナル強度を16~64倍も向上させることができます。

高いレベルでのタンパク質発現

エピトープ・タグの付加機能の他に、pCMV-3Tag Vectorは哺乳類細胞において高レベルでのタンパク質発現を実現します。クローニングされた遺伝子は、サイト・メガロウイルス(CMV)プロモーターにより、幅広い哺乳類細胞で恒常的に発現されます。また、N-末端ベクターを使用の際は、Kozak(コザック)コンセンサス配列により融合タンパク質の最適な発現をもたらします。高い発現レベルと柔軟なエピトープ・タグの付加機能により、pCMV-3Tag Vectorシリーズは、目的のタンパク質にタグを付加し解析する際の最適なツールになります。

柔軟な製品構成

FLAG®/c-MycとN-末端/C-末端の組み合わせが、それぞれ2種類のベクターであり、合計で8種類の製品を用意しています。pCMV- 3Tag-1~4は、導入された細胞へネオマイシン/カナマイシン耐性を付与し、それぞれ1塩基ずつリーディング・フレームが異なる3つのベクターを1セットとしています。3つのリーディング・フレームにより、目的の遺伝子が発現のフレームを合わせてエピトープ・タグへ融合することを可能にします。pCMV-3Tag-6~9は、安定した形質導入体の選択ができるように、ハイグロマイシン耐性遺伝子が組み込まれています。

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We compared activity levels of the luciferase protein fused to one and three epitope tags.
Our pCMV-3Tag vector series provides maximum versatility in tagging orientation, multiple reading frames, and antibiotic resistance.
Western blot analysis of serial dilutions of the luciferase protein expressed with one or three copies of the FLAG® or c-Myc epitope tags. Triple epitope-tagged proteins are detected with up to 60 fold higher sensitivity than single epitope tagged proteins. Note triple tagged luciferase has a higher molecular weight than single tagged. Similar signal comparisons were seen with N-terminus tags (data not shown).

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>>  Licensing

The use of the CMV Promoter is covered under U.S. Patent Nos. 5,168,062 and 5,385,839 owned by the University of Iowa Research Foundation and licensed FOR RESEARCH USE ONLY. For further information, please contact UIRF at 319-335-4546.
FLAG® License Agreement
The enclosed DNA expression vector and/or antibody are specifically adpated for a method of producing selected protein molecules covered by one or more of the following patents owned by Sigma-Aldrich Co.: U.S. Patent Nos. (5,011912, 4,703,004, 4,782,137 and 4,851,341;EP Patent No. 150,126 (Austria, Belgium, Switzerland, France, United Kingdom, Italy, Netherlands and Sweden); EP Patent No. 335,899 (Belgium, Switzerland, Germany, France, United Kingdom, Italy, Luxembourg and Sweden); German Patent No. P3584260.1; Canadian Patent No. 1,307,752; and Japanese Patent Nos. 1,983,150 and 2,665,359. Your payment includes a limited license under these
patents to make only the following uses of these products:
A. Vector License: You may use the enclosed vector to transform cells to produce proteins containing the amino acid sequence DYKDDDDK for research purposes provided, however, such research purposes do not include binding an unlicensed antibody to any portion of this amino acid sequence nor using such proteins for the preparation of antibodies having an affinity for any portion of this amino acid sequence.
B. Antibody License: You may only use the enclosed antibody for research purposes to perform a method of producing a protein in which the protein is expressed in a host cell and purified by use of the antibody in accordance with a claim in one of the above patents in force in a country where the use actually occurs so long as: (1) you perform such method with a DNA expression vector licensed from Sigma-Aldrich Co.; and (2) you do not bind (or allow others to bind) an unlicensed antibody to any DYKDDDDK epitope of any fusion protein that is produced by use of the method.
This license does not include any rights under any other patents. You are not licensed to use the vector and/or antibody in any manner or for any purposed not recited above. As used above, the term 'unlicensed antibody” means any antibody which Sigma-Aldrich Co. has not expressly licensed pursuant to Paragraph B, above. Sigma-Aldrich Co. hereby expressly retains all rights in the above listed patents not expressly licensed hereunder.
If the terms and conditions of this License Agreement are acceptable to you, then you may open the vessel(s) containing the vector and/or antibody and, through such act of opening a vessel, will have shown your acceptance to these terms and conditions. If the terms and conditions of this License Agreement are not acceptable to you, then please return the vessel(s) unopened to Agilent for a complete refund of your payment.
For additional licensing information or to receive a copy of any of the above patents, please contact the Sigma-Aldrich Co. licensing department at telephone number 314-771-5765.

 
Product Catalog # Amount Price

pCMV-3Tag-1 Epitope Tagging Mammalian Expression Vectors

240195 3 x 20 µg お問い合わせください Call
pCMV-3Tag-1 vectors fuse three copies of FLAG epitope tag to the N-terminus of proteins expressed in mammalian cells for stronger signal in your protein characterization experiments. Neomycin/Kanamycin resistance.
N-terminal FLAG tagging vectors in all 3 reading frames
分析保証書(C of A) 分析保証書 (C of A)   マニュアル マニュアル

pCMV-3Tag-2 Epitope Tagging Mammalian Expression Vectors

240196 3 x 20 µg お問い合わせください Call
pCMV-3Tag-2 vectors fuse three copies of c-Myc epitope tag to the N-terminus of proteins expressed in mammalian cells for stronger signal in your protein characterization experiments. Neomycin/Kanamycin resistance.
N-terminal c-Myc tagging vectors in all 3 reading frames
分析保証書(C of A) 分析保証書 (C of A)   マニュアル マニュアル

pCMV-3Tag-3 Epitope Tagging Mammalian Expression Vectors

240197 3 x 20 µg お問い合わせください Call
pCMV-3Tag-3 vectors fuse three copies of FLAG epitope tag to the C-terminus of proteins expressed in mammalian cells for stronger signal in your protein characterization experiments. Neomycin/Kanamycin resistance.
C-terminal FLAG tagging vectors in all 3 reading frames
分析保証書(C of A) 分析保証書 (C of A)   マニュアル マニュアル

pCMV-3Tag-4 Epitope Tagging Mammalian Expression Vectors

240198 3 x 20 µg お問い合わせください Call
pCMV-3Tag-4 vectors fuse three copies of c-Myc epitope tag to the C-terminus of proteins expressed in mammalian cells for stronger signal in your protein characterization experiments. Neomycin/Kanamycin resistance.
C-terminal c-Myc tagging vectors in all 3 reading frames
分析保証書(C of A) 分析保証書 (C of A)   マニュアル マニュアル

pCMV-3Tag-6 vectors

240200 20 µg お問い合わせください Call
pCMV-3Tag-6 vectors fuse three copies of FLAG epitope tag to the N-terminus of proteins expressed in mammalian cells for stronger signal in your protein characterization experiments. Hygromycin resistance.
N-terminal FLAG tagging vectors
分析保証書(C of A) 分析保証書 (C of A)   マニュアル マニュアル

pCMV-3Tag-7 vectors

240202 20 µg お問い合わせください Call
pCMV-3Tag-7 vectors fuse three copies of c-Myc epitope tag to the N-terminus of proteins expressed in mammalian cells for stronger signal in your protein characterization experiments. Hygromycin resistance.
N-terminal c-Myc tagging vectors
分析保証書(C of A) 分析保証書 (C of A)   マニュアル マニュアル

pCMV-3Tag-8 vectors

240203 20 µg お問い合わせください Call
pCMV-3Tag-8 vectors fuse three copies of FLAG epitope tag to the C-terminus of proteins expressed in mammalian cells for stronger signal in your protein characterization experiments. Hygromycin resistance.
C-terminal FLAG tagging vectors
分析保証書(C of A) 分析保証書 (C of A)   マニュアル マニュアル

pCMV-3Tag-9 vectors

240204 20 µg お問い合わせください Call
pCMV-3Tag-9 vectors fuse three copies of c-Myc epitope tag to the C-terminus of proteins expressed in mammalian cells for stronger signal in your protein characterization experiments. Hygromycin resistance.
C-terminal c-Myc tagging vectors
分析保証書(C of A) 分析保証書 (C of A)   マニュアル マニュアル