NuPAGE™ Bis-Tris Welcome Pack, 10%

Name: NuPAGE™ Bis-Tris Welcome Pack, 10%
SKU: NP030A

El paquete de bienvenida NuPAGE Bis-Tris proporciona todos los geles, tampones y reactivos NuPAGE que necesita para empezar a usar el depósito de minigel.

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Número de catálogo	Pocillos
NP030A	10 pocillo
NP030B	12 pocillo
NP030C	15 pocillo

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Número de catálogo NP030A

Precio (MXN)

Pocillos:

10 pocillo

El paquete de bienvenida NuPAGE Bis-Tris proporciona todos los geles, tampones y reactivos NuPAGE que necesita para empezar a usar el depósito de minigel. El depósito de minigel es compatible con todos los minigeles Invitrogen Novex, NuPAGE y Bolt. Cada depósito de minigel puede albergar hasta dos geles por ciclo. El exclusivo diseño del depósito permite cargar cómodamente el gel en paralelo y mejora la visualización durante el uso. Los tiempos de procesamiento pueden variar en función de las condiciones del tampón y las fuentes de alimentación que se empleen.

El paquete de bienvenida Bis-Tris NuPAGE contiene:
• Depósito de minigel (A25977)
• Minigeles de bis-tris NuPAGE (dos cajas, 20 geles)
• Tampón de desplazamiento de MES SDS NuPAGE (20X) (NP0002)
• Tampón de muestra de LDS NuPAGE (4X) (NP0007)
• Agente reductor de muestras NuPAGE (10X) (NP0009)
• Escalera de proteínas preteñida PageRuler Plus, de 10 a 250 kDa (26619)

Acerca del depósito de minigel
El depósito de minigel es compatible con todos los minigeles Invitrogen Novex, NuPAGE y Bolt. Cada depósito de minigel puede albergar hasta dos geles por ciclo. El exclusivo diseño del depósito permite cargar cómodamente el gel en paralelo y mejora la visualización durante el uso. Los tiempos de procesamiento pueden variar en función de las condiciones del tampón y las fuentes de alimentación que se empleen.

Acerca de los geles NuPAGE Bis-Tris
Los geles de proteínas Invitrogen NuPAGE Bis-Tris son geles de poliacrilamida prefundidos diseñados para una separación óptima de una amplia gama de proteínas en condiciones desnaturalizantes. A diferencia de los geles tradicionales de Tris-glicina, los geles NuPAGE Bis-Tris tienen un ambiente de pH neutro que minimiza las modificaciones de proteínas. Utilice los geles NuPAGE Bis-Tris para preparar las proteínas para la secuenciación, la espectrometría de masas y cualquier otra técnica donde la integridad de las proteínas resulte crucial. Además, utilice los geles NuPAGE para obtener resultados óptimos durante el día a día.

Transferencia
Para transferir proteínas a una membrana, recomendamos utilizar el tampón de transferencia NuPAGE (NP0006) para la transferencia húmeda tradicional utilizando el módulo Mini Blot (B1000). También se puede realizar una transferencia semiseca rápida con Invitrogen Power Blotter o una transferencia rápida en seco con el dispositivo de transferencia de gel iBlot 2 (IB21001).

For Research Use Only. Not for use in diagnostic procedures.

Especificaciones

Gel Thickness1,0 mm

Línea de productosNuPAGE

Cantidad1 paquete de bienvenida

Condiciones de envíoAprobado para su envío a temperatura ambiente y en hielo húmedo

Para utilizar con (equipo)Depósito de minigel

Porcentaje del gel10 %

Tamaño de gelMini

Tipo de gelBis-Tris

Intervalo de separaciónDe 3,5 a 260 kDa

Pocillos10 pocillo

Unit SizeEach

Contenido y almacenamiento

• Depósito de minigel
• Geles de proteína Bis-Tris NuPAGE al 10 %, 1,0 mm, 10 pocillos
• Tampón de desplazamiento MES SDS NuPAGE (20X)
• Tampón de muestra LDS NuPAGE (4X)
• Agente reductor de muestras NuPAGE (10X)
• Estándar de proteínas preteñidas PageRuler Plus, de 10 a 250 kDa

Preguntas frecuentes

I used one of your protein standards for a western transfer and noticed that some of the lower-molecular weight protein bands passed through the membrane. How can I resolve this issue?

- Decrease voltage, current or length of transfer time
- Make sure that the methanol concentration in the transfer buffer is proper; use a methanol concentration of 10-20% methanol removes the SDS from SDS-protein complexes and improves the binding of protein to the membrane.
- Make sure that the SDS concentration (if added) in the transfer buffer is proper, don't use more than 0.02-0.04% SDS. Using too much SDS can prevent binding of proteins to the membrane.
- Check the pore size of the membrane and the size of the target protein. Proteins smaller than 10 kDa will easily pass through a 0.45 µm pore size membrane. If proteins smaller than 10 kDa are of interest, it would be better to use a 0.2 µm pore size membrane.

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

I used one of your protein standards for a western transfer and noticed that some of the higher-molecular weight bands transferred very poorly to the membrane. Can you offer some tips?

- Increase voltage, current or length of time for transfer
- SDS in the gel and in the SDS-protein complexes promotes elution of the protein from the gels but inhibits binding of the protein to membranes. This inhibition is higher for nitrocellulose than for PVDF. For proteins that are difficult to elute from the gel such as large molecular weight proteins, a small amount of SDS may be added to the transfer buffer to improve transfer. We recommend pre-equilibrating the gel in 2X Transfer buffer (without methanol) containing 0.02-0.04% SDS for 10 minutes before assembling the sandwich and then transferring using 1X transfer buffer containing 10% methanol and 0.01%SDS.
- Methanol removes the SDS from SDS-protein complexes and improves the binding of protein to the membrane, but has some negative effects on the gel itself, leading to a decrease in transfer efficiency. It may cause a reduction in pore size, precipitation of some proteins, and some basic proteins to become positively charged or neutral. Make sure that the methanol concentration in the transfer buffer is not more than 10-20% and that high-quality, analytical grade methanol is used.

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

I used one of your pre-stained standards on a Tris-Glycine gel and noticed that the molecular weights of the proteins were different than on a NuPAGE Bis-Tris gel. What is the reason for this?

Pre-stained standards have a dye that is covalently bound to each protein that will result in the standard migrating differently in different buffer systems (i.e., different gels). As a result, using a pre-stained standard for molecular weight estimation will only give the apparent molecular weight of the protein. Pre-stained standards may be used for molecular weight approximation, confirming gel migration and estimating blotting efficiency but for accurate molecular weight estimation, an unstained standard should be used.

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

I used one of your protein standards and am seeing some extra bands in the lane. Can you offer some suggestions?

- While loading, take care to make sure that there is no cross-contamination from adjacent sample lanes.
- Make sure that the correct amount of standard is loaded per lane. Loading too much protein can result in extra bands and this is a problem especially with silver-stained gels.
- Improper storage of the standard or repeated freeze/thawing can result in protein degradation.

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

I used one of your protein standards and the bands look non-distinct and smeary. What should I do?

Here are some suggestions:

- Make sure that the correct amount of standard is loaded per lane. Loading too much protein can cause smearing and this is a problem especially with silver stained gels.
- Bands will not be as well resolved in low percentage gels. Try using a higher percentage gel.
- If the bands look smeary and non-distinct after a western transfer/detection, this may be due to the antibody being too concentrated. Follow the manufacturer's recommended dilution or determine the optimal antibody concentration by dot-blotting.

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

View all NuPAGE™ Bis-Tris Welcome Pack, 10% FAQs