Enteroquinasa EKMax™
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Invitrogen™

Enteroquinasa EKMax™

EKMax™ es una preparación recombinante de la subunidad catalítica de la enteroquinasa bovina (1). EKMax™ reconoce la secuencia Asp-Asp-Asp-Asp-Lys yMás información
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Número de catálogoCantidad
E180021000 unidades
E18001250 unidades
Número de catálogo E18002
Precio (USD)
-
Cantidad:
1000 unidades
EKMax™ es una preparación recombinante de la subunidad catalítica de la enteroquinasa bovina (1). EKMax™ reconoce la secuencia Asp-Asp-Asp-Asp-Lys y escinde el enlace péptido después del residuo de lisina. La enzima se puede utilizar para escindir cualquier proteína de fusión que lleve esta secuencia de péptidos (Figura 1).

Aplicación: Eliminación de etiquetas de fusión de proteínas recombinantes.

Definición de la unidad: Una unidad de EKMax™ es la cantidad de enzima necesaria para digerir 20 µg de proteína de fusión de tiorredoxina-CAT al 90 % de su finalización en 16 horas a 37 °C. Una unidad de EKMax™ es equivalente a ∼190 unidades de activación de tripsinógeno.
Para uso exclusivo en investigación. No apto para uso en procedimientos diagnósticos.
Especificaciones
Tipo de productoEnteroquinasa
Cantidad1000 unidades
EspecieGanado/bovino
Línea de productosEKMax™
Unit SizeEach
Contenido y almacenamiento
La enteroquinasa EKMax™ se suministra con tampón 10X. Almacenar a -20 °C. Estabilidad garantizada durante seis meses si se almacena correctamente.

Preguntas frecuentes

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.

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