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Invitrogen™

Vivid Colors™ pcDNA™6.2/C-EmGFP-GW/TOPO™ Mammalian Expression Vector

Los vectores de expresión TOPO™ de proteínas fluorescentes Vivid Colors™ pcDNA™6.2 (Figura 1) le permiten clonar rápidamente un gen yMás información
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Número de catálogoCantidad
K3592020 reacciones
Número de catálogo K35920
Precio (USD)
-
Cantidad:
20 reacciones
Los vectores de expresión TOPO™ de proteínas fluorescentes Vivid Colors™ pcDNA™6.2 (Figura 1) le permiten clonar rápidamente un gen y fusionarlo con las proteínas fluorescentes (FPS) ampliamente utilizadas y bien caracterizadas de la medusa «Aequorea victoria» (1, 2). Los potentes vectores de clonación TOPO™ contienen proteína fluorescente verde esmeralda (EmGFP) o proteína fluorescente amarilla (YFP) para una detección simple y no invasiva de la proteína recombinante (Figura 2). Ambas proteínas fluorescentes se han humanizado para dar una expresión óptima en mamíferos (3). Los vectores de expresión TOPO™ de proteínas fluorescentes Vivid Colors™ pcDNA™6.2 ofrecen:
• Topoisomerasa I para la clonación TOPO™ de un paso y 5 minutos del gen de interés amplificado por PCR
• Promotor de CMV para la expresión de alto nivel de la proteína de fusión fluorescente recombinante
• Capacidad para fusionar EmGFP o YFP al terminal N o C de una proteína
• Marcador de resistencia Bsd para la selección rápida de líneas celulares estables
Para uso exclusivo en investigación. No apto para uso en procedimientos diagnósticos.
Especificaciones
Sistema constitutivo o inducibleConstitutivo
Tipo de entregaTransfección
Para utilizar con (aplicación)Ensayo con indicadores, localización subcelular
Tipo de productoVector de expresión de mamíferos
Cantidad20 reacciones
Gen marcadorGFP (EmGFP)
Agente de selección (eucariótico)Blasticidina
VectorpcDNA
Método de clonaciónTOPO-TA
Línea de productosGateway, TOPO, Vivid Colors, pcDNA
PromotorCMV
Etiqueta de proteínaGFP (EmGFP)
Unit SizeEach
Contenido y almacenamiento
Cada kit de vector de expresión pcDNA™6.2-TOPO™ contiene dos cajas. La caja TOPO™ contiene vector pcDN™6.2-TOPO™ linealizado y activado por topoisomerasa I, plantilla de control, cebadores para secuenciación, componentes para la reacción de PCR y un plásmido de control de expresión. La caja TOPO™ debe conservarse a -20°C. La caja One Shot ™ contiene reactivos de transformación que incluyen alícuotas de un solo uso de 50 μl de E. coli One Shot™ TOP10 químicamente competente, medio S.O.C. y un plasmídico de control superenrollado pUC19. Conservar la E. coli competente a -80°C. Se garantiza la estabilidad de todos los reactivos durante 6 meses si se almacenan correctamente.

Preguntas frecuentes

Your Gateway-adapted TOPO vectors are supplied with a control template and control primers. Can I obtain the sequence of the control template?

The sequence of the control template is proprietary.

Can GFP fluorescence be detected in cells that have been stained for beta-galactosidase?

We recommend looking for GFP fluorescence before staining for beta-galactosidase. This is because the beta-galactosidase staining process produces a very high autofluorescence that will interfere with detection of GFP fluorescence.

What are the recommended filter sets for detection of EmGFP, YFP, CFP, and BFP by fluorescence microscopy?

EmGFP, YFP, CFP, and BFP can be detected using standard FITC filter sets and settings. However, for optimal detection of the fluorescence signal, filter sets optimized for detection within the excitation and emission ranges for each fluorescent protein are recommended. The recommended filter sets are as follows: EmGFP: Omega filter set XF100 YFP: Omega filter set XF1042 Chroma filter set 41028 CFP: Omega filter set XF114 Chroma filter set 31044 BFP: Omega filter set XF10 Chroma filter set 31021 For information on obtaining filter sets, please contact Omega Optical, Inc. (www.omegafilters.com) or Chroma Technology Corporation (www.chroma.com) directly.

What are the excitation and emission maxima for your fluorescent proteins (EmGFP, YFP, BFP, CFP, and Cycle 3 GFP)?

Excitation and emission maxima for our fluorescent proteins are as follows:
- EmGFP: Excitation: 487 nm; Emission: 509 nm
- YFP: Excitation: 514 nm; Emission: 527 nm
- BFP: Excitation: 308-383 nm; Emission: 440-447 nm
- CFP: Excitation: 452 nm; Emission: 505 nm
- Cycle 3 GFP: Primary excitation: 395 nm; Secondary Excitation: 478 nm; Emission: 507 nm

Are the fluorescent proteins you offer (EmGFP, YFP, CFP, BFP, and Cycle 3 GFP) humanized?

Yes, all of the fluorescent proteins offered by us (EmGFP, YFP, CFP, BFP, and Cycle 3 GFP) have been humanized for optimal mammalian expression.

Citations & References (4)

Citations & References
Abstract
Ultra-high-throughput screening method for the directed evolution of glucose oxidase.
Authors:Ostafe R, Prodanovic R, Nazor J, Fischer R,
Journal:
PubMed ID:24613019
'Glucose oxidase (GOx) is used in many industrial processes that could benefit from improved versions of the enzyme. Some improvements like higher activity under physiological conditions and thermal stability could be useful for GOx applications in biosensors and biofuel cells. Directed evolution is one of the currently available methods to ... More
Development of a fluorescence-based method for monitoring glucose catabolism and its potential use in a biomass hydrolysis assay.
Authors:Haney LJ, Coors JG, Lorenz AJ, Raman DR, Anex RP, Scott MP,
Journal:Biotechnol Biofuels
PubMed ID:19019221
'The availability and low cost of lignocellulosic biomass has caused tremendous interest in the bioconversion of this feedstock into liquid fuels. One measure of the economic viability of the bioconversion process is the ease with which a particular feedstock is hydrolyzed and fermented. Because monitoring the analytes in hydrolysis and ... More
Lignin depletion enhances the digestibility of cellulose in cultured xylem cells.
Authors:Lacayo CI, Hwang MS, Ding SY, Thelen MP,
Journal:
PubMed ID:23874568
'Plant lignocellulose constitutes an abundant and sustainable source of polysaccharides that can be converted into biofuels. However, the enzymatic digestion of native plant cell walls is inefficient, presenting a considerable barrier to cost-effective biofuel production. In addition to the insolubility of cellulose and hemicellulose, the tight association of lignin with ... More
Surface carbohydrate analysis and bioethanol production of sugarcane bagasse pretreated with the white rot fungus, Ceriporiopsis subvermispora and microwave hydrothermolysis.
Authors:Sasaki C, Takada R, Watanabe T, Honda Y, Karita S, Nakamura Y, Watanabe T,
Journal:Bioresour Technol
PubMed ID:21903385
'Effects of pretreatments with a white rot fungus, Ceriporiopsis subvermispora, and microwave hydrothermolysis of bagasse on enzymatic saccharification and fermentation were evaluated. The best sugar yield, 44.9 g per 100g of bagasse was obtained by fungal treatments followed by microwave hydrothermolysis at 180°C for 20 min. Fluorescent-labeled carbohydrate-binding modules which ... More