pEXP1-DEST Vector Kit

Citas y referencias (2)

Invitrogen™

pEXP1-DEST Vector Kit

Para adaptarse a todas sus necesidades de expresión, Invitrogen ofrece vectores de destino Gateway™ de vanguardia de destino para laMás información

Número de catálogo	Cantidad
V96001	6 μg

Número de catálogo V96001

Precio (MXN)

Cantidad:

Pedido a granel o personalizado

Para adaptarse a todas sus necesidades de expresión, Invitrogen ofrece vectores de destino Gateway™ de vanguardia de destino para la expresión en célula de E. coli, insecto, levadura o mamífero, así como para la producción de proteína nativa o proteínas de fusión N o C-terminal. Los vectores de destino Gateway™ tienen sitios attR para la recombinación con cualquier fragmento flanqueado por attL, independientemente de si se trata de un clon de entrada o de un clon Ultimate™ RF. La siguiente tabla enumera la amplia gama de vectores de destino disponibles.

Materiales adicionales necesarios, disponibles por separado: Clon de entrada Gateway™, mezcla de enzimas Gateway™ LR Clonase™ y tampón de reacción.

Para uso exclusivo en investigación. No apto para uso en procedimientos diagnósticos.

Especificaciones

Resistencia bacteriana a los antibióticosAmpicilina (AMPR)

HendiduraSitio de reconocimiento de EK (enterocinasa)

Tipo de productoVector de expresión de destino del sistema Gateway

Cantidad6 μg

Agente de selección (eucariótico)Ninguno

VectorpEXP, pDEST

Método de clonaciónGateway

Línea de productosExpressway, Gateway

PromotorT7

Etiqueta de proteínaEtiqueta His (6x)

Unit Size6 µg

Contenido y almacenamiento

Todos los vectores de destino se suministran liofilizados y superenrollados.

Preguntas frecuentes

I accidentally stored my E. coli slyD-Extract, E. coli Reaction Buffer (-A.A.), and 2X feed buffer at room temperature. Can I still use them?

Unfortunately, this may result in a loss of activity.

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

I've run out of the T7 RNA polymerase for my cell-free expression. What do you suggest I use?

We would recommend using T7 RNA polymerase (Cat. No. 18033019, 50 U/µL). Use 1-1.5 µL in a 50 µL reaction system.

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

I'm getting smearing after running my cell-free expression reaction on a gel.. What could be the cause of this?

Smearing may occur if samples for the following reasons:

- Samples were not precipitated with acetone: precipitate proteins with acetone to remove background smearing.
- Too much protein was loaded: reduce the amount used.
- The gel itself was not clean: rinse the gel briefly before exposing to film.
- Ethanol was present in the protein synthesis reaction: make sure that any residual ethanol is removed during DNA purification.
- Check the date of your pre-cast gels: do not use gels after the expiration date.

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

I'm seeing a ladder of small-sized products after running my reaction on a gel when using the Expressway system. Why is this?

There may be several reasons for why this is occurring. The most common are: proteolysis, degradation of DNA and/or RNA templates (truncated templates will generate truncated protein products), internal initiation (if there are many methionines and internal RBS-like sequences in the gene, the ribosome may initiate translation from the wrong methionine), premature termination, translational pausing, frequent rare codon usage, complicated secondary structure of RNA, and others. This can also happen if proteins are denatured for too long, or not enough SDS was added to the 1X SDS-PAGE sample buffer.

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

With a cell-free expression system, I'm getting good protein yield, but it has low biological activity. What can I do?

- Your protein may not be folding properly: try to reduce the incubation temperature to as low as 25 degrees C during synthesis.
- You may require post-translational modification of your protein: the Expressway system will not introduce post-translational modifications to the recombinant protein.
- Your synthetic protein may require co-factors for complete activity: try adding required co-factors to the protein synthesis reaction.

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

View all pEXP1-DEST Vector Kit FAQs

Citations & References (2)

Citations & References

Abstract

BimEL is an important determinant for induction of anoikis sensitivity by mitogen-activated protein/extracellular signal-regulated kinase kinase inhibitors.

Authors:Fukazawa H, Noguchi K, Masumi A, Murakami Y, Uehara Y,

Journal:Mol Cancer Ther

PubMed ID:15486195

'Loss of contact with substratum triggers apoptosis in many normal cell types, a phenomenon termed anoikis. We reported previously that mitogen-activated protein/extracellular signal-regulated kinase kinase (MEK) inhibitors induced apoptosis in nonanchored MDA-MB231 and HBC4 human breast cancer cells, whereas anchored cells remained viable. Here, we report that activation of the ... More

Fox-2 splicing factor binds to a conserved intron motif to promote inclusion of protein 4.1R alternative exon 16.

Authors:Ponthier JL, Schluepen C, Chen W, Lersch RA, Gee SL, Hou VC, Lo AJ, Short SA, Chasis JA, Winkelmann JC, Conboy JG,

Journal:J Biol Chem

PubMed ID:16537540

'Activation of protein 4.1R exon 16 (E16) inclusion during erythropoiesis represents a physiologically important splicing switch that increases 4.1R affinity for spectrin and actin. Previous studies showed that negative regulation of E16 splicing is mediated by the binding of heterogeneous nuclear ribonucleoprotein (hnRNP) A/B proteins to silencer elements in the ... More