Estándar de proteínas de inmunotransferencia MagicMark™ XP
Estándar de proteínas de inmunotransferencia MagicMark™ XP
Estándar de proteínas de inmunotransferencia MagicMark™ XP
Estándar de proteínas de inmunotransferencia MagicMark™ XP
Invitrogen™

Estándar de proteínas de inmunotransferencia MagicMark™ XP

El estándar de proteínas de inmunotransferencia MagicMark XP se compone de nueve proteínas recombinantes (20–220 kDa), cada una de lasMás información
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Número de catálogoCantidad
LC560350 μl
LC5602250 μl
Número de catálogo LC5603
Precio (USD)
230,04
Each
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Cantidad:
50 μl
Pedido a granel o personalizado
Precio (USD)
230,04
Each
Añadir al carro de la compra
El estándar de proteínas de inmunotransferencia MagicMark XP se compone de nueve proteínas recombinantes (20–220 kDa), cada una de las cuales contiene un sitio de unión de IgG. El sitio de unión de IgG enlaza el anticuerpo primario o secundario utilizado para la detección de la proteína objetivo, lo que permite una visualización directa del estándar en la inmunotransferencia (Western Blot). El estándar MagicMark XP es compatible con la mayoría de los kits y sustratos de inmunotransferencia (quimioluminiscentes, cromogénicos y fluorescentes). El estándar de proteínas se suministra en un formato listo para usar para la carga directa en geles; sin necesidad de calentar, reducir o añadir el tampón de muestra antes de su uso.

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

Aplicaciones
• Western blotting: de las nueve bandas sin teñir mediante el método de detección utilizado para la proteína diana. Compatible con sustratos quimioluminiscentes y anticuerpos secundarios fluorescentes (no recomendado para anticuerpos marcados con fluoruros en el canal de 500–550 nm).
• Estimación del tamaño de las proteínas: medición de las proteínas en geles de poliacrilamida-SDS y en Western blot a través del método de detección Western Blot
Para uso exclusivo en investigación. No apto para uso en procedimientos diagnósticos.
Especificaciones
Método de detecciónSistema de detección definido por el usuario
Compatibilidad del gelGeles Novex™ tricina, geles Novex™ Tris-glicina , geles NuPAGE™ Bis-Tris, geles NuPAGE™ Tris-acetato, geles SDS-PAGE
Peso molecular220, 120, 100, 80, 60, 50, 40, 30, 20 kDa
Línea de productosMagicMark
Tipo de productoMarcadores moleculares de proteínas
Cantidad50 μl
Listo para cargarSí
Condiciones de envíoHielo seco
Tipo de tinciónSin teñir
Tipo de sistemaWestern Blotting, SDS-PAGE
Number of Markers9
Intervalo de tamañosDe 20 a 220 kDa
Unit SizeEach
Contenido y almacenamiento
50 μl de mezcla estándar lista para usar suministrada en un vial de plástico. El tampón de carga consta de 125 mM de Tris-HCl (pH 6,8), 10 mM de DTT, glicerol al 17,4 %, SDS al 3 % y bromofenol azul al 0,025 %.

Almacenar a -20 °C.

Preguntas frecuentes

I used your MagicMark XP Western Protein Standard and obtained a very weak or almost no signal after the western detection. Can you please help me troubleshoot?

Here are some suggestions:

- Verify that the detection reagents are working well. Optimize the antibody concentration to obtain best results.
- Make sure that the amount of standard loaded on the gel is correct.
- Optimize the transfer conditions (current, voltage, transfer time).
- Enzyme-conjugated primary antibodies may not bind efficiently with the proteins in the MagicMark XP Western Protein Standard. We recommend using unconjugated primary antibody, followed by the addition of enzyme-conjugated secondary antibody.
Note: The Anti-myc-AP/HRP and Anti-V5-AP/HRP antibodies do not bind to MagicMark XP proteins.

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.

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.

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