Estándar de proteínas de amplia gama sin teñir PageRuler™
Thermo Scientific™
Estándar de proteínas de amplia gama sin teñir PageRuler™
La escalera de proteínas de rango amplio sin teñir Thermo Scientific PageRuler es una mezcla de 11 proteínas (de 5Más información
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Número de catálogo
Cantidad
26630
2 x 250 μl
Número de catálogo 26630
Precio (MXN)
-
Cantidad:
2 x 250 μl
Pedido a granel o personalizado
La escalera de proteínas de rango amplio sin teñir Thermo Scientific PageRuler es una mezcla de 11 proteínas (de 5 a 250 kDa) para su uso como patrones de tamaño en electroforesis de proteínas (SDS-PAGE) y Western blots. La escalera 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.
Características del producto • Bandas de referencia: las bandas de 100, 50 y 20 kDa son más intensas para facilitar la orientación • Etiquetado: cada proteína de la escalera contiene una secuencia integral Strep-tag™ II y se puede detectar directamente en Western blots mediante conjugados de Strep-Tactin™ o un anticuerpo frente a la secuencia Strep-tag™ II.
Aplicaciones • Dimensionamiento preciso de proteínas en geles de poliacrilamida-SDS y Western blots
Para uso exclusivo en investigación. No apto para uso en procedimientos diagnósticos.
Especificaciones
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
Tipo de productoMarcadores moleculares de proteínas
Cantidad2 x 250 μl
Listo para cargarSí
Condiciones de envíoAprobado para su envío en hielo húmedo o seco
Tipo de tinciónSin teñir
Tipo de sistemaWestern Blotting, SDS-PAGE
Number of Markers11
Intervalo de tamañosDe 5 a 250 kDa
Unit SizeEach
Contenido y almacenamiento
Contenido: dos viales de 250 μL cada uno
Tampón de almacenamiento: 62,5 mM Tris-H3PO4 (pH 7,5 a 25 °C), 1 mM EDTA, 2 % SDS, 10 mM DTT, 1 mM NaN3, 0,01 % azul de bromofenol y 33 % glicerol
almacenamiento: Tras su recepción, almacenar a -20 °C
Preguntas frecuentes
The upper bands of the ladder are missing. What could be the reason?
The upper bands of the ladder may be degraded by proteases. Ladder, gel, buffer, pipettes, pipette tips, or equipment can be contaminated by proteases during usage. A general recommendation would be to avoid working with proteases in the same room. We would recommend preparing fresh solutions, cleaning the equipment, and using clean pipettes and tips. If the ladder itself is contaminated, please use a new tube of the ladder.
Some additional bands or smears were observed on the gel when using a PageRuler unstained ladder. What may have caused this?
Additional bands can appear due to dithiothreitol (DTT) oxidation in the storage buffer. Please add newly prepared DTT solution to the final concentration of 100 mM and boil for 5 min at 95 degrees C. This should solve the issue. Addition of DTT is NOT recommended for prestained protein ladders, since too high a concentration of reducing agents can cause protein destaining.
Do the proteins in Thermo Scientific protein ladders have a His-Tag or would otherwise react with an anti-His-Tag antibody?
No, proteins in Thermo Scientific protein ladders are not His tagged. However, non-specific interaction between the ladder proteins and primary or secondary antibodies is possible and some His-Tag detection systems, such as Thermo Scientific 6xHis Protein Tag Stain Reagent Kit, show non-specific interaction. The protein ladder bands are more readily detected when using high antibody concentrations. The non-specific cross-reactivity is difficult to predict, it often has a different pattern dependent on the antibodies used in each individual experiment. The most general way to handle this problem would be to use lower concentrations of antibodies and to use lower amount of protein ladders. It may also be useful to leave one empty well between the ladder and the sample to overcome a possible leakage of the signal to the nearby sample lane.
Can Thermo Scientific protein ladders be detected by Strep-Tactin conjugates?
PageRuler Unstained protein ladders can be detected directly on Western blots by using Strep-Tactin conjugates or an antibody against the Strep-tag II sequence. All PageRuler and Spectra ladder proteins contain an integral Strep-tag II sequence, however the prestained ladders cannot be detected by Strep-Tactin conjugates.
Why is non-specific binding detected after Western blot?
Protein ladder bands can sometimes be detected with chemiluminescent techniques due to non-specific interactions of ladder proteins with either primary or secondary antibodies (or with both). The ladder bands are only rarely detected by chromogenic substrates. The extremely high sensitivity of the chemiluminescent assays is needed to see the bands, so the actual degree of cross-reactivity is low. The non-specific cross-reactivity is difficult to predict, it often has a different pattern depending on the antibodies used. If antibodies recognize a linear epitope, the cross-reactivity may be due to sequence homology. If antibodies react with a denaturation-resistant conformational epitope it could be impossible to identify the exact reason for detected cross-reactivity. The most general way to handle this problem would be to use lower concentrations of antibodies.