HiMark™ Unstained Protein Standard

El estándar de proteínas de alto peso molecular (HMW) sin teñir HiMark contiene nueve proteínas de 40–500 kDa diseñadas paraMás información

Número de catálogo	Cantidad
LC5688	250 μl

Número de catálogo LC5688

Precio (USD)

Cantidad:

250 μl

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El estándar de proteínas de alto peso molecular (HMW) sin teñir HiMark contiene nueve proteínas de 40–500 kDa diseñadas para el análisis de proteínas de alto peso molecular. 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.

Compare y visualice todos los demás estándares y escaleras de proteínas ›

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 preteñidas de alto peso molecular HiMark también se puede utilizar con los geles NuPAGE Novex Bis-Tris al 4–12 % y el tampón de desplazamiento NuPAGE MOPS y los geles de Tris-glicina Novex al 4 %.

Aplicaciones
• Dimensionamiento preciso de proteínas de alto peso molecular en geles de poliacrilamida-SDS y Western blots

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

Especificaciones

Para utilizar con (aplicación)Tris-acetate gels

Compatibilidad del gelGeles Bolt™ Bis-Tris Plus, Geles de tricina Novex™, geles de Tris-glicina Novex™, geles NuPAGE™ Bis-Tris, geles de Tris-acetato NuPAGE™, geles SDS-PAGE

Peso molecular500, 290, 240, 160, 116, 97, 66, 55, 40 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 tinciónSin teñir

Tipo de sistemaWestern Blotting, SDS-PAGE

Number of Markers9

Intervalo de tamañosDe 40 a 500 kDa

Unit SizeEach

Contenido y almacenamiento

Contenido: un vial de 250 μl

Tampón de almacenamiento: 250 Mm Tris-HCl (pH 8,5), 0,5 mm EDTA, 50 mm DTT, 10 % glicerol, 2 % LDS, 0,2 mm Coomassie G-250y 0,175 mm Phenol Red

almacenamiento: Tras su recepción, almacenar a 20 °C

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 HiMark™ Unstained Protein Standard FAQs