EKMax™ Enterokinase
Product Image
Citations & References (4)
Invitrogen™

EKMax™ Enterokinase

EKMax™はウシエンテロキナーゼ（1）の触媒サブユニットの組換え調製です。EKMax™は配列Asp-Asp-Asp-Asp-Lysを認識し、リジン残基の後のペプチド結合を切断します。酵素は、このペプチド配列を保持しているあらゆる融合タンパク質を切断するために使用できます（図1）。用途：組換えタンパク質からの融合タグの除去詳細を見る
テクニカルサポートオーダーサポート
表示を変更buttonViewtableView
製品番号（カタログ番号）数量
E18001250ユニット
E180021000ユニット
製品番号（カタログ番号） E18001
価格（JPY）
132,200
Each
お問い合わせください ›
数量:
250ユニット
EKMax™はウシエンテロキナーゼ（1）の触媒サブユニットの組換え調製です。EKMax™は配列Asp-Asp-Asp-Asp-Lysを認識し、リジン残基の後のペプチド結合を切断します。酵素は、このペプチド配列を保持しているあらゆる融合タンパク質を切断するために使用できます（図1）。

用途：組換えタンパク質からの融合タグの除去。

単位の定義：EKMax™の1つの単位は、20µgのチオレドキシン-CAT融合タンパク質を37℃で16時間で90%消化するために必要な酵素の量です。EKMax™の1ユニットは、約190トリプシンゲン活性化ユニットに相当します。
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
製品タイプエンテロキナーゼ
数量250ユニット
種畜牛／ウシ
製品ラインEKMax
Unit SizeEach
組成および保存条件
1000または250ユニットのEKMax™を使用すると、バッファーが10倍になります。-20℃で保存適切に保存されている場合、6ケ月間安定していることが保証されます。

よくあるご質問（FAQ）

What is the molecular weight of the EKMax enterokinase enzyme?

EKMax enterokinase is a clone of the catalytic subunit of enterokinase expressed in the yeast Pichia pastoris. The calculated molecular weight of the protein is 26.3 kDa, but it contains three sites for asparagine-linked glycosylation. The apparent molecular weight of 43 kDa is consistent with previous observations (LaVallie et al., 1993) and is assumed to be because of N-linked glycosylation.
Reference: LaVallie, E.R., Rehemtulla, A., Racie, L.A.,Diblasio, E.A., Ferenz, C., Grant, K.L. Light, A., and McCoy, J.M. (1993). Cloning and functional expression of a cDNA encoding the catalytic subunit of bovine enterokinase. J.Biol.Chem. 268, 23311-23317.

Will DTT, Triton X-100 detergent, Tween 20 detergent, Thesit, calcium chloride, sodium chloride, or SDS affect the efficiency of Enterokinase (EKMax) enzyme cleavage?

Enterokinase is active in buffers containing up to 1 mM DTT, 0.1% Triton X-100 detergent, 0.1% Tween 20 detergent, and 0.1% Thesit. It is recommended to have 10mM Tris pH 8.0 and 10 mM calcium chloride in the buffer. Enterokinase is inhibited by sodium chloride and SDS.

Should Enterokinase be resuspended in a buffer containing 50% glycerol to protect the protein from freeze/thaw cycles?

Freeze thaw has a minimal effect on the activity of Enterokinase. The addition of glycerol is not necessary but can make handling of the enzyme easier.

How specific is cleavage by EKMax Enterokinase? Are there any alternate cleavage sites for the enzyme?

Enterokinase cleaves after the sequence (Asp)4-Lys.

It has been proposed that the active center of enterokinase possesses a distinctive cationic subsite that binds -(Asp)4. Enterokinase is highly specific and tolerates very few changes to its recognition site. If the ionic charge of the recognition site is preserved, enterokinase will recognize the site, but the rate of hydrolysis of the peptide bond will be reduced (Light and Janska, 1989). The four aspartyl residues act as a signal for enterokinase cleavage. It has been reported that with only three aspartyl residues the rate of hydrolysis is reduced. Two aspartyl residues preceding the lysyl residue are the minimum number of acidic residues needed to maintain specificity (Maroux et al., 1971). Non-specific cleavage by enterokinase may occur in the cases described above, but this is usually alleviated by reducing the amount of enzyme used.

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 (4)

引用および参考文献
Abstract
Interaction of the Conserved Region 4.2 of sigma E with the RseA Anti-sigma Factor.
Authors: Tam Christina; Collinet Bruno; Lau Gary; Raina Satish; Missiakas Dominique;
Journal:J Biol Chem
PubMed ID:12016219
Evarsigma(E) RNA polymerase transcribes a regulon of folding factors for the bacterial envelope and is induced by physical and chemical stresses. The RseA anti-sigma factor inhibits the activity of Evarsigma(E) RNA polymerase. It is shown here that the N-terminal portion of varsigma(E), residues 1-153, binds core RNA polymerase. RseA interacts ... More
Heme oxygenase-2 is a hemoprotein and binds heme through heme regulatory motifs that are not involved in heme catalysis.
Authors:McCoubrey WK Jr, Huang TJ, Maines MD
Journal:J Biol Chem
PubMed ID:9139709
The heme oxygenase (HO) system degrades heme to biliverdin and CO and releases chelated iron. In the primary sequence of the constitutive form, HO-2, there are three potential heme binding sites: two heme regulatory motifs (HRMs) with the absolutely conserved Cys-Pro pair, and a conserved 24-residue heme catalytic pocket with ... More
Identification of amino acid residues critical for LD78beta, a variant of human macrophage inflammatory protein-1alpha, binding to CCR5 and inhibition of R5 human immunodeficiency virus type 1 replication.
Authors: Miyakawa Toshikazu; Obaru Kenshi; Maeda Kenji; Harada Shigeyoshi; Mitsuya Hiroaki;
Journal:J Biol Chem
PubMed ID:11734558
In an attempt to determine which amino acid(s) of LD78beta, a variant of human macrophage inflammatory protein-1alpha, plays a critical role in the interaction with CCR5, we generated six LD78beta variants with an amino acid substituted to Ala at the NH(2) terminus of LD78beta. There was no significant difference in ... More
Spinesin/TMPRSS5, a novel transmembrane serine protease, cloned from human spinal cord.
Authors: Yamaguchi Nozomi; Okui Akira; Yamada Tatsuo; Nakazato Hiroshi; Mitsui Shinichi;
Journal:J Biol Chem
PubMed ID:11741986
A cDNA encoding a novel serine protease, which we designated spinesin, has been cloned from human spinal cord. The longest open reading frame was 457 amino acids. A homology search revealed that the human spinesin gene was located at chromosome 11q23 and contained 13 exons, the gene structure being similar ... More