Estándar de proteínas fluorescentes Benchmark™

El patrón de proteínas fluorescente BenchMark consta de una amplia gama de peso molecular de proteínas (∼11–155 kDa) conjugadas conMás información

Número de catálogo	Cantidad
LC5928	125 μL

Número de catálogo LC5928

Precio (MXN)

Cantidad:

125 μL

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El patrón de proteínas fluorescente BenchMark consta de una amplia gama de peso molecular de proteínas (∼11–155 kDa) conjugadas con el colorante fluorescente Alexa Fluor 488, lo que permite la visualización directa mediante fluorescencia. Las proteínas se pueden resolver en bandas nítidas mediante SDS-PAGE y visualizar mediante un transiluminador UV, una cámara CCD con los filtros adecuados o sistemas de escáner láser.

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Características del producto
• Visualización: fluorescencia en el canal de 488 nm, excitación máxima a 493 nm y emisión máxima a 516 nm
• Formato listo para usar: 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.
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 de poliacrilamida-SDS fluorescentes y Western blots

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

Especificaciones

Método de detecciónCanal de 488 nm

Compatibilidad del gelGeles de Tris-Glicina Novex™, geles de NuPAGE™ Bis-Tris

Peso molecular155, 98, 63, 40, 32, 21, 11 kDa

Línea de productosBenchMark

Tipo de productoMarcadores moleculares de proteínas

Cantidad125 μL

Listo para cargarSí

Condiciones de envíoHielo seco

Tipo de tinciónBandas conjugadas con Alexa Fluor 488

Tipo de sistemaWestern Blotting, SDS-PAGE

Number of Markers7

Intervalo de tamañosDe 11 a 155 kDa

Unit SizeEach

Contenido y almacenamiento

Se proporcionan 125 μL (25-50 aplicaciones) en tampón de carga que consta de Tris-HCl, SDS, glicerol y azul de coomassie™ G-250.

Almacenar a -20 °C. Evite repetir la congelación y descongelación Se garantiza la estabilidad durante 6 meses si se almacena correctamente.

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.

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