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pcDNA™3.1 (+) Mammalian Expression Vector
pcDNA&trade;3.1<sup> (+)</sup> Mammalian Expression Vector
Invitrogen™

pcDNA™3.1 (+) Mammalian Expression Vector

このpcDNA™3.1(+)ベクターは、さまざまな哺乳類細胞株における高レベルでの構成的発現のために設計されています。Geneticin™選択可能マーカーおよびフォワード方向の複数のクローニング部位を含みます。pcDNA™3.1発現ベクターファミリーpcDNA™3.1の3つのタグなしバージョン(個別に使用可能)には、それぞれに異なる選択可能マーカー(Geneticin™、ゼオシン™、またはHygromycin詳細を見る
製品番号(カタログ番号)数量
V7902020 μg
製品番号(カタログ番号) V79020
価格(JPY)
103,200
20 µg
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数量:
20 μg
Ask our AI about this Product
このpcDNA™3.1(+)ベクターは、さまざまな哺乳類細胞株における高レベルでの構成的発現のために設計されています。Geneticin™選択可能マーカーおよびフォワード方向の複数のクローニング部位を含みます。

pcDNA™3.1発現ベクターファミリー
pcDNA™3.1の3つのタグなしバージョン(個別に使用可能)には、それぞれに異なる選択可能マーカー(Geneticin™、ゼオシン™、またはHygromycin)があり、単独で使用することも、相互コトランスフェクションで使用することもできます。3つのベクターは以下の特長を備えています:

•高レベル発現用のサイトメガロウイルス(CMV)エンハンサープロモーター
• フォワード(+)方向の大規模な複数のクローニング部位
• ウシ成長ホルモン(BGH)ポリアデニル化シグナルおよびmRNA安定性向上のための転写終了配列
• ラージT抗原(COS-1やCOS-7など)を発現する細胞株内でのエピソーム複製および単純なベクターレスキュー用のSV40起源
• アンピシリン耐性遺伝子および大腸菌の選択および維持のためのpUC起源
研究用にのみ使用できます。診断用には使用いただけません。
仕様
供給タイプTransfection
使用対象(アプリケーション)構成的発現
製品タイプ哺乳類発現用ベクター
数量20 μg
選択剤(真核生物)Geneticin™(G-418)
ベクターpcDNA
クローニング法制限酵素/MCS
製品ラインpcDNA™
プロモーターCMV
タンパク質タグタグなし
Unit Size20 µg
組成および保存条件
20 µgのこのpcDNA™3.1(+)ベクターと発現コントロールが、超らせん型で凍結乾燥された状態で提供されます。-20℃で保存ベクターは、適切に保存されている場合、6カ月間安定していることが保証されます。
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Safety Data Sheets

ベクター情報

ベクターマップ
マップ
ポリリンカー
配列
制限
pcDNA4/TO/Myc-His
pcDNA3.1
pcDNA3/CAT (replaced with pcDNA3.1/CAT)
pcDNA3.1(+)

よくあるご質問(FAQ)

Here are possible causes and solutions:

Detection method may not be appropriate or sensitive enough:
- We recommend optimizing the detection protocol or finding more sensitive methods. If the protein is being detected by Coomassie/silver staining, we recommend doing a western blot for increased sensitivity. The presence of endogenous proteins in the lysate may obscure the protein of interest in a Coomassie/silver stain. If available, we recommend using a positive control for the western blot.
- Insufficient number of clones screened: Screen at least 20 clones.
- Inappropriate antibiotic concentration used for stable selection: Make sure the antibiotic kill curve was performed correctly. Since the potency of a given antibiotic depends upon cell type, serum, medium, and culture technique, the dose must be determined each time a stable selection is performed. Even the stable cell lines we offer may be more or less sensitive to the dose we recommend if the medium or serum is significantly different.
- Expression of gene product (even low level) may not be compatible with growth of the cell line: Use an inducible expression system.
- Negative clones may result from preferential linearization at a vector site critical for expression of the gene of interest: Linearize the vector at a site that is not critical for expression, such as within the bacterial resistance marker.

Here are possible causes and solutions:

- Try the control expression that is included in the kit
Possible detection problem:

- Detection of expressed protein may not be possible in a transient transfection, since the transfection efficiency may be too low for detection by methods that assess the entire transfected population. We recommend optimizing the transfection efficiency, doing stable selection, or using methods that permit examination of individual cells. You can also increase the level of expression by changing the promoter or cell type.
- Expression within the cell may be too low for the chosen detection method. We recommend optimizing the detection protocol or finding more sensitive methods. If the protein is being detected by Coomassie/silver staining, we recommend doing a western blot for increased sensitivity. The presence of endogenous proteins in the lysate may obscure the protein of interest in a Coomassie/silver stain. If available, we recommend using a positive control for the western blot. Protein might be degraded or truncated: Check on a Northern. Possible time-course issue: Since the expression of a protein over time will depend upon the nature of the protein, we always recommend doing a time course for expression. A pilot time-course assay will help to determine the optimal window for expression. Possible cloning issues: Verify clones by restriction digestion and/or sequencing.

Find additional tips, troubleshooting help, and resources within our Protein Expression Support Center.

No; neomycin is toxic to mammalian cells. We recommend using Geneticin (a.k.a. G418 Sulfate), as it is a less toxic and very effective alternative for selection in mammalian cells.

Translation initiation will occur at the first ATG encountered by the ribosome, although in the absence of a Kozak sequence, initiation will be relatively weak. Any insert downstream would express a fusion protein if it is in frame with this initial ATG, but levels of expressed protein are predicted to be low if there is a non-Kozak consensus sequence. If the vector contains a non-Kozak consensus ATG, we recommend that you clone your gene upstream of that ATG and include a Kozak sequence for optimal expression.

pcDNA3.1 vectors contain the core CMV promoter that is truncated before the start of transcription, whereas the pcDNA 3.3-TOPO vector has the 672 bp native CMV promoter. This native CMV promoter allows high-level gene expression with two- to five-fold higher protein yields compared to other expression vectors. pcDNA3.1 vectors are available in restriction, TOPO, and Gateway cloning versions and as untagged and epitope-tagged versions, whereas the pcDNA3.3-TOPO vector is a TOPO TA-adapted, untagged vector that can be used to express native proteins without extraneous amino acids, and is hence ideal for antibody production and structural biology.

引用および参考文献 (340)

引用および参考文献
Abstract
Two murine homologs of the Drosophila single-minded protein that interact with the mouse aryl hydrocarbon receptor nuclear translocator protein.
Authors:Probst MR,Fan CM,Tessier-Lavigne M,Hankinson O
Journal:The Journal of biological chemistry
PubMed ID:9020169
Evaluation of VP22 spread in tissue culture.
Authors:Brewis N,Phelan A,Webb J,Drew J,Elliott G,O'Hare P
Journal:Journal of virology
PubMed ID:10623773
We compare methods of detection of intercellular transport of the herpes simplex virus protein VP22 and of a green fluorescent protein (GFP)-VP22 fusion protein. Spread of both proteins was observed by immunofluorescence (IF) using organic fixatives. Spread of both proteins was also detected by IF after paraformaldehyde (PFA) fixation and ... More
A kinase-regulated PDZ-domain interaction controls endocytic sorting of the beta2-adrenergic receptor.
Authors:Cao TT,Deacon HW,Reczek D,Bretscher A,von Zastrow M
Journal:Nature
PubMed ID:10499588
Molecular cloning and biological activity of a novel lysyl oxidase-related gene expressed in cartilage.
Authors:Ito H; Akiyama H; Iguchi H; Iyama K; Miyamoto M; Ohsawa K; Nakamura T;
Journal:J Biol Chem
PubMed ID:11292829
We cloned a cDNA encoding a novel lysyl oxidase-related protein, named LOXC, by suppression subtractive hybridization between differentiated and calcified ATDC5 cells, a clonal mouse chondrogenic EC cell line. The deduced amino acid sequence of mouse LOXC consists of 757 amino acids and shows 50% identity with that of mouse ... More
A
Authors:Chen Zhenhui; Alcayaga Carmen; Suarez-Isla Benjamin A; O'Rourke Brian; Tomaselli Gordon; Marban Eduardo;
Journal:J Biol Chem
PubMed ID:11973330
The large size (six membrane-spanning repeats in each of four domains) and asymmetric architecture of the voltage-dependent Na+ channel has hindered determination of its structure. With the goal of determining the minimum structure of the Na+ channel permeation pathway, we created two stable cell lines expressing the voltage-dependent rat skeletal ... More
340 total citations

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