Sistema de expresión Expi293™ MembranePro™

Name: Sistema de expresión Expi293™ MembranePro™
SKU: A25870

Para los investigadores que estudian los receptores G acoplados a proteínas (GPCR) y otras proteínas de membrana, el sistema deMás información

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Número de catálogo	Cantidad
A25870	100 reacciones
A25869	10 reacciones

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Número de catálogo A25870

Precio (USD)

Cantidad:

100 reacciones

Para los investigadores que estudian los receptores G acoplados a proteínas (GPCR) y otras proteínas de membrana, el sistema de expresión Expi293™ MembranePro™ ofrece proteínas de membrana funcionales enriquecidas de alta eficiencia, escalabilidad y fiabilidad para una amplia variedad de aplicaciones de bajo o alto rendimiento.

Las características del sistema de expresión Expi293™ MembranePro™ incluyen:
•Eficiencia: aumento en más de 20 veces de la producción de proteínas de membrana en comparación con el sistema de cultivo adherente
• Escalable: fácil de escalar y sin necesidad de múltiples matraces
• Facilidad de uso: no se requiere disociación enzimática o mecánica

Además, los beneficios de la tecnología MembranePro™ incluyen:
•Las proteínas se muestran en las membranas celulares humanas (o de otros mamíferos)
• El control de calidad celular y el procesamiento postraslacional de mamíferos ayudan a producir proteínas funcionales
• Las proteínas brotan como lipopartículas y se recogen fácilmente del medio del cultivo
• Las lipopartículas se enriquecen con receptores expresados

Con el protocolo del sistema de expresión Expi293™ MembranePro™, que requiere menos trabajo, podrá:
1.Clonar su gen de interés mediante un vector TOPO™
2.Transfectar un cultivo en suspensión de células Expi293F™ utilizando el reactivo de transfección Expifectamine™ 293
3.Cosechar lipopartículas 48 h después de la adición de potenciadores utilizando la mezcla de precipitación MembranePro™
4.Almacenar lipopartículas a -80 ºC o proceder a los ensayos posteriores

Nota:Se requieren células Exi293F™ (A14527) y un kit de expresión pEF6 V5 TOPO™ (K9610-20) para el uso de este sistema y deben adquirirse por separado cuando sea necesario.

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

Especificaciones

Línea de célulasExpi293F™

Tipo de entregaPartículas similares a virus (VLP)

Para utilizar con (aplicación)Expresión de proteínas de membrana

Línea de productosExpi293, MembranePro

Cantidad100 reacciones

Suficiente para100 reacciones

PromotorEF-1α

TipoSistema de expresión

Unit SizeEach

Contenido y almacenamiento

• MembranePro™ Reagent, 10 x 0.3 mg (1 mg/mL), store at -5 to -30°C
• MembranePro™ Precipitation Mix, 10 x 75 mL, store at room temperature
• 10 ExpiFectamine™ 293 transfection kits (containing reagent and enhancers) each for 1 L of culture, store at 2–8°C
• Opti-MEM™ I Reduced Serum Medium, 10 x 100 mL, store at 2–8°C
• Expi293™ Expression Medium, 10 x 1 L, store in the dark at 2–8°C

Preguntas frecuentes

The Expi293F cells supplied with the Expi293 MembranePro Expression System had poor viability upon thawing. Can I use a different cell line?

While other cell lines can be tested, we recommend using the Expi293F cells, as the performance of the kit has been optimized for use with this cell line (i.e., transfection efficiency using ExpiFectamine Transfection Reagent).

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

Can I use a different expression vector with the MembranePro Functional Protein Expression Kit or the Expi293 MembranePro Expression System?

While other expression vectors and promoters can be tested, we recommend using the pEF6 V5-His TOPO TA expression vector for optimal yields, as the performance of these kits has been optimized for use with this expression vector containing the EF-1alpha promoter.

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

Is there a positive control included with the MembranePro Functional Protein Expression Kit or the Expi293 MembranePro Expression System?

No. However, you can visually tell if MembranePro particles formed via a pellet at the bottom of the tube following precipitation. You can also test the function of your MembranePro particles via receptor-ligand binding studies.

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

I am interested in expressing some functional membrane proteins in mammalian cells. Do you offer a system that will allow me to do this?

We offer the MembranePro Functional Protein Expression System for the expression and display of mammalian cell surface membrane proteins, including G-protein coupled receptors (GPCRs), in an aqueous-soluble format. The system uses virus-like particles (VLPs) to capture lipid raft regions of the cell’s plasma membrane as the VLPs are secreted from the cell. Using this system, it is possible to capture and display endogenous or overexpressed GPCRs and other cell surface membrane proteins in their native context for downstream assays. Since the VLPs are packaged by the cell and secreted into the culture medium, VLPs allow for the isolation of functional membrane proteins by simply decanting and clarifying the culture medium, and isolating the VLPs by precipitation. This represents a substantial savings in time, effort, and required machinery over preparing cell membrane fractions. Because VLPs capture receptor-rich regions of the plasma membrane, your GPCR may also be substantially enriched over crude membrane preparations.

The standard MembranePro Functional Protein Expression System is offered in two configurations: The MembranePro Functional Protein Expression Kit (Cat. No. A11667), which provides the convenience of a complete kit with all the reagents supplied for 10 reactions, and the MembranePro Functional Protein Support Kit, which includes the reagents for functional expression of membrane proteins but does not contain the expression vector cloning module or the 293FT host cells. The MembranePro Functional Protein Support Kit is offered in three sizes, allowing 10, 60, or 600 reactions (Cat. Nos. A11668, A11669, A11670, respectively).

For high-yield expression of functional membrane proteins in Exp293F cells, we offer the Expi293 MembranePro Expression System (Cat. Nos. A25869, A25870) that combines the scalability and ease of use of Expi293 and the technology of MembranePro to allow an increase of more than 20-fold in membrane protein yield compared to the standard, adherent culture MembranePro Functional Protein Expression System. Please see the Application Note for more details.

Note: Expi293F cells (Cat. No. A14527) and pEF6 V5 His TOPO Expression Vector Kit (Cat. No. K961020) are not supplied with the Expi293 MembranePro Expression System and have to be purchased separately as required.

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

I have misplaced the pEF6 V5-His TOPO TA expression vector from the Expi293 MembranePro Expression System. Can I use a different expression vector?

While other expression vectors and promoters can be tested, we recommend using the pEF6 V5-His TOPO TA expression vector for optimal yields, as the performance of the kit has been optimized for use with this expression vector containing the EF-1alpha promoter.

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

View all Sistema de expresión Expi293™ MembranePro™ FAQs