Marcadores de peso molecular de proteínas preteñidos HiMark™

El estándar de proteínas preteñido HiMark está diseñado para el análisis de proteínas de alto peso molecular y consta deMás información

Número de catálogo	Cantidad
LC5699	250 μl

Número de catálogo LC5699

Precio (MXN)

Cantidad:

250 μl

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El estándar de proteínas preteñido HiMark está diseñado para el análisis de proteínas de alto peso molecular y consta de nueve bandas de proteínas de 30–460 kDa. El estándar de proteínas se suministra en un formato listo para su uso para la carga directa en geles; sin necesidad de calentar, reducir ni añadir un tampón de muestras antes de usarlo.

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Para obtener los mejores resultados y la separación de proteínas de alto peso molecular, le recomendamos el uso de geles de Tris-acetato. El estándar de proteínas de alto peso molecular preteñido HiMark también puede usarse con geles NuPAGE Bis-Tris al 4–12 % y tampón de desplazamiento NuPAGE MOPS y con geles de Tris-glicina Novex al 4 %.

Aplicaciones
• Supervisión de la migración de proteínas durante la electroforesis en gel de poliacrilamida-SDS
• Supervisión de la transferencia de proteínas a las membranas tras Western blot
• Dimensionamiento de proteínas en geles SDS-PAGE y Western blots

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

Especificaciones

Método de detecciónColorimétrico

Para utilizar con (aplicación)Tris-acetate gels

Compatibilidad del gelGeles de Tris-glicina Novex™, geles de Tris-acetato NuPAGE™

Peso molecular460, 268, 238, 171, 117, 71, 55, 41, 31 kDa

Línea de productosHiMark

Tipo de productoMarcadores moleculares de proteínas

Cantidad250 μl

Listo para cargarSí

Aplicaciones recomendadasGeles de Tris-acetato

Condiciones de envíoHielo seco

Tipo de tinción2 colores: Morado, rosa

Tipo de sistemaWestern Blotting, SDS-PAGE

Number of Markers9

Intervalo de tamañosDe 30 a 460 kDa

Unit SizeEach

Contenido y almacenamiento

Contenido: 250 μL de patrón de proteínas de alto peso molecular (HMW) preteñido HiMark™
Tampón de almacenamiento: Tris-HCl, formamida, SDS, rojo de fenol
Almacenamiento: almacenar a -20 °C

Preguntas frecuentes

Can I use the HiMark High Molecular Weight Protein Standard with your NuPAGE Bis-Tris or NuPAGE Tris-Glycine gels?

Yes, the HiMark High Molecular Weight Protein Standard can be used with our NuPAGE gels, though we would recommend using our NuPAGE Tris-Acetate gels for best results when separating high moleuclar weight proteins.

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

I used one of your pre-stained protein standards for a western transfer and I noticed that the intensity of the band faded from the membrane during the transfer process. Why is this?

The fading is most likely due to detergent in the western blocking/washing solutions that can remove some of the proteins from the membrane. The dye itself will not wash off of the proteins because it is covalently bound. We have found that smaller pore size membranes retain the proteins better during blocking and wash procedures, and hence recommend use of 0.2 µm instead of 0.45 µm membranes for best resolution and protein retention. After transfer, it is a good idea to circle the pre-stained bands with a pencil on the membrane, so band positions can be identified after blocking and processing.

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 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.

View all Marcadores de peso molecular de proteínas preteñidos HiMark™ FAQs