WesternBreeze™ Chromogenic Kit, anti-mouse

Los kits cromogénicos WesternBreeze® detectan proteínas que han sido inmovilizadas en membranas (nitrocelulosa o PVDF) después de la transferencia WesternMás información

Número de catálogo	Cantidad
WB7103	1 kit

Número de catálogo WB7103

Precio (MXN)

Cantidad:

1 kit

Los kits cromogénicos WesternBreeze® detectan proteínas que han sido inmovilizadas en membranas (nitrocelulosa o PVDF) después de la transferencia Western o enlazadas directamente desde la solución (Dot Blot). La detección se realiza con un sustrato BCIP/NBT listo para usar para fosfatasa alcalina. Los precipitados de color morado que no se desvanecen se desarrollan en las bandas de proteínas de la membrana. WesternBreeze® (cromogénicos) ofrece:

• Fondo transparente
• Niveles bajos de picogramos de alta sensibilidad detectables
• Alta especificidad
• Un protocolo simple, sin necesidad de optimización

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

Especificaciones

Reactividad cruzadaRatón

Cantidad1 kit

ReactividadRatón

Condiciones de envíoHielo húmedo

Tipo de sustratoSustrato de AP (fosfatasa alcalina)

Método de detecciónCromogénico

Membrane CompatibilityNitrocelulosa, PVDF

Línea de productosWesternBreeze

Tipo de productoKit western blot

Unit Size1 kit

Contenido y almacenamiento

Los kits cromogénicos WesternBreeze™ incluyen soluciones de bloqueo, diluyente de anticuerpos primarios, solución de anticuerpos secundarios lista para usar (antirratón, anticonejo o anticaprino), sustrato cromogénico listo para usar, soluciones de lavado y bandejas de incubación. Cada kit contiene reactivos completos para 20 miniblots. Almacenar los kits a 4 °C. Se garantiza la estabilidad durante 6 meses si se almacena correctamente.

Preguntas frecuentes

Why is the actual band size on a western blot different from the predicted size of the protein?

Western blotting is based on the separation of proteins by their size on a gel. However, migration of proteins through the gel matrix is also affected by other factors, which may cause the observed band size to be different from the predicted size.

Common causes are:
-Post-translational modification; for example phosphorylation and glycosylation increase the size of the protein
-Post-translation cleavage; many proteins are synthesized as precursor proteins, and then cleaved to give the active form
-Multimers, for example dimerization of a protein. This is usually prevented under reducing conditions, although strong interactions can result in the appearance of higher bands
-Splice variants; alternative splicing may result in different sized proteins being produced from the same gene
-Relative charge; the composition of amino acids (charged vs. non-charged)

Find additional tips, troubleshooting help, and resources within our Protein Electrophoresis and Western Blotting Support Center.

What are the standard lysis buffers used with mammalian cells for detection of protein expression by immunoprecipitation (IP) or Western blot analysis?

The most commonly used buffer is RIPA Buffer with SDS. We offer RIPA Buffer (Cat. Nos. 89900 and 89901). We also offer the Pierce IP Lysis buffer (Cat. Nos. 87787 and 87788) as well as M-PER (Cat. Nos. 78501, 78503, and 78505).

Find additional tips, troubleshooting help, and resources within our Cell Lysis and Fractionation Support Center.

What conditions do you recommend for overnight Western transfer?

Doing an overnight Westerm transfer is not the preferred method but can be done. The power should be lowered and the buffer should be chilled or the unit should be placed in the cold room to prevent overheating. You may try an overnight transfer at 5-15 V and adjust accordingly. You may also wish to put a second membrane behind the first in order to bind any proteins that transfer through the first membrane. You can use both membranes for staining, immunoblotting, or analysis.

Find additional tips, troubleshooting help, and resources within our Protein Electrophoresis and Western Blotting Support Center.

After transferring onto a PVDF membrane, what is the best way to store the membrane for future probing?

We recommend air drying the PVDF membrane and placing it in an envelope, preferably on top of a supported surface to keep the membrane flat. It can be stored indefinitely at ≤80 degrees C. Right before probing, we recommend re-wetting the membrane with alcohol for a few seconds, followed by a few rinses with pure water to reduce the alcohol concentration. Then proceed as normal with the blocking step.

Find additional tips, troubleshooting help, and resources within our Protein Electrophoresis and Western Blotting Support Center.

Which transfer buffers are recommended for peptide (N-terminal) sequencing?

Use non-glycine based buffers such as the NuPAGE Invitrogen Transfer buffer, CAPS, or 1/2X TBE transfer buffer.

Find additional tips, troubleshooting help, and resources within our Protein Electrophoresis and Western Blotting Support Center.

View all WesternBreeze™ Chromogenic Kit, anti-mouse FAQs

Citations & References (1)

Citations & References

Abstract

Molecular and functional analysis of the lepB gene, encoding a type I signal peptidase from Rickettsia rickettsii and Rickettsia typhi.

Authors:Rahman MS, Simser JA, Macaluso KR, Azad AF,

Journal:J Bacteriol

PubMed ID:12867468

'The type I signal peptidase lepB genes from Rickettsia rickettsii and Rickettsia typhi, the etiologic agents of Rocky Mountain spotted fever and murine typhus, respectively, were cloned and characterized. Sequence analysis of the cloned lepB genes from R. rickettsii and R. typhi shows open reading frames of 801 and 795 ... More