AcTEV™ Protease

Citations & References (6)

Invitrogen™

AcTEV™ Protease

Name: AcTEV™ Protease
SKU: 12575015

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製品番号（カタログ番号）	数量
12575015	1000 U
12575023	10,000 U

2 オプション

製品番号（カタログ番号） 12575015

価格（JPY）

55,600

Each

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数量:

1000 U

AcTEV™プロテアーゼは、7アミノ酸の配列（Glu-Asn-Leu-Tyr-Phe-Gln-Gly）を認識し、この配列中のGlnとGlyの間を特異的に切断します。これは融合タンパク質からアフィニティータグを除去する場合に役立ちます。AcTEV™プロテアーゼは、高い部位特異性と活性を備え、ネイティブなTobacco Etch Virus（TEV）プロテアーゼよりも大幅に安定性を向上させたTEVプロテアーゼの改良版です。この改良により結果的に活性を長期に保つことができます。AcTEV™プロテアーゼの特長：

•特異性の高い切断活性
• 長期にプロテアーゼ活性を維持するために酵素の安定性を向上させています（図を参照）
• 幅広い温度（+4℃～30℃）およびpH（6.0～8.5）において活性があります
• 6-His配列を利用して、消化されたタンパク質サンプルから取り除くことができます
• 非特異的なプロテアーゼの混入がなく、シングルバンドで85%以上の純度があります

アプリケーション
AcTEV™プロテアーゼによるインキュベーションは、融合タグから目的のタンパク質を放出。これは、組み換えタンパク質から、溶解、分泌、検出および精製タグを除去する効果的な方法です。

酵素の仕様
AcTEV™プロテアーゼ遺伝子を発現する大腸菌から精製。

ユニット定義
AcTEV™プロテアーゼの1ユニットで、30℃の条件下、1時間で3 µgのコントロール基質の85%が切断されます。

ユニット反応条件
30 µL中に50 mMのトリス-HCl（pH 8.0）、0.5 mMのEDTA、1 mMのDTT、3 µgのコントロール基質、および1ユニットの酵素を加え、30℃で1時間反応。AcTEV™プロアテーゼは、非特異的なプロアテーゼ活性がないことを機能的に試験しています。

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

適合バッファーTEV Buffer

製品タイプProtease

数量1000 U

出荷条件ドライアイス

濃度1,000 Units

酵素TEV Protease

最適反応温度+4°C to +30°C

製品ラインAcTEV

Unit SizeEach

組成および保存条件

AcTEV™ Proteaseには、20X TEVバッファー[1 M Tris-HCl（pH 8.0）、10 mM EDTA]のバイアル 1本と100 mM DTTのバイアル1本が付属しています。

-20℃で保存。適切に保存した場合、1年間安定しています。

よくあるご質問（FAQ）

Why does AcTEV Protease require DTT?

A final concentration of 1 mM DTT is required for the AcTEV protease reaction. The DTT serves as a stabilizer of secondary structure, i.e., it ensures that the enzyme does not undergo oxidation. If you are unable to use DTT due to a column purification after digestion, you can leave it out and still see a successful AcTEV digestion, however we recommend using it.

No rigorous quantitative data for activity in the absence of DTT and EDTA has been collected in-house. Incubations using reaction buffer +/-DTT and +/- EDTA; have been performed, but the products were analyzed only at t=0 and t=2 hr at 30 degrees C. No difference in the amount of product generated +/-DTT or +/-EDTA was noticed, but it is possible that the detailed kinetics in a time course reaction may be a little different.

What is the cleavage recognition site for AcTEV Protease?

The recognition site for AcTEV Protease is ENLYFQS/G

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

What products do you offer for enzymatic cleavage of fusion tags from recombinant proteins?

We offer the following products:

-AcTEV Protease (Cat. Nos. 12575015, 12575023)
-EKMax Enterokinase (Cat. Nos. E18001, E18002)
-SUMO Protease (Cat. No. 12588018)

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

引用および参考文献 (6)

引用および参考文献

Abstract

Structure and function of an irreversible agonist-ß(2) adrenoceptor complex.

Authors:Rosenbaum DM, Zhang C, Lyons JA, Holl R, Aragao D, Arlow DH, Rasmussen SG, Choi HJ, Devree BT, Sunahara RK, Chae PS, Gellman SH, Dror RO, Shaw DE, Weis WI, Caffrey M, Gmeiner P, Kobilka BK,

Journal:Nature

PubMed ID:21228876

'G-protein-coupled receptors (GPCRs) are eukaryotic integral membrane proteins that modulate biological function by initiating cellular signalling in response to chemically diverse agonists. Despite recent progress in the structural biology of GPCRs, the molecular basis for agonist binding and allosteric modulation of these proteins is poorly understood. Structural knowledge of agonist-bound ... More

Quantitative reactivity profiling predicts functional cysteines in proteomes.

Authors:Weerapana E, Wang C, Simon GM, Richter F, Khare S, Dillon MB, Bachovchin DA, Mowen K, Baker D, Cravatt BF,

Journal:Nature

PubMed ID:21085121

'Cysteine is the most intrinsically nucleophilic amino acid in proteins, where its reactivity is tuned to perform diverse biochemical functions. The absence of a consensus sequence that defines functional cysteines in proteins has hindered their discovery and characterization. Here we describe a proteomics method to profile quantitatively the intrinsic reactivity ... More

Widespread bidirectional promoters are the major source of cryptic transcripts in yeast.

Authors:Neil H, Malabat C, d'Aubenton-Carafa Y, Xu Z, Steinmetz LM, Jacquier A,

Journal:Nature

PubMed ID:19169244

'Pervasive and hidden transcription is widespread in eukaryotes, but its global level, the mechanisms from which it originates and its functional significance are unclear. Cryptic unstable transcripts (CUTs) were recently described as a principal class of RNA polymerase II transcripts in Saccharomyces cerevisiae. These transcripts are targeted for degradation immediately ... More

Non-muscle myosin IIA is a functional entry receptor for herpes simplex virus-1.

Authors:Arii J, Goto H, Suenaga T, Oyama M, Kozuka-Hata H, Imai T, Minowa A, Akashi H, Arase H, Kawaoka Y, Kawaguchi Y,

Journal:Nature

PubMed ID:20944748

Herpes simplex virus-1 (HSV-1), the prototype of the a-herpesvirus family, causes life-long infections in humans. Although generally associated with various mucocutaneous diseases, HSV-1 is also involved in lethal encephalitis. HSV-1 entry into host cells requires cellular receptors for both envelope glycoproteins B (gB) and D (gD). However, the gB receptors ... More

Hexameric assembly of the proteasomal ATPases is templated through their C termini.

Authors:Park S, Roelofs J, Kim W, Robert J, Schmidt M, Gygi SP, Finley D,

Journal:Nature

PubMed ID:19412160

Substrates of the proteasome are recognized and unfolded by the regulatory particle, and then translocated into the core particle (CP) to be degraded. A hetero-hexameric ATPase ring, containing subunits Rpt1-6, is situated within the base subassembly of the regulatory particle. The ATPase ring sits atop the CP, with the Rpt ... More

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