Coomassie Gel Staining | Consejo Superior de Investigaciones Científicas

Service data

Support unit: PROTEÓMICA
Institute: CENTRO DE BIOLOGIA MOLECULAR SEVERO OCHOA
Locality: Madrid (Madrid)
Service's web: http://www.cbm.csic.es

Service's description

La investigación Proteómica implica la identificación y la caracterización de todo el perfil proteico de una célula, tejido u organismo, en un contexto espacial y temporal. El estudio incluye isoformas, mutantes, modificaciones e interacciones proteína-proteína/s. La trayectoria del Servicio de Proteómica ha sido paralela a la evolución de la comunidad científica, y sus objetivos son ofrecer soporte científico-técnico al investigador, con métodos apropiados para la preparación de la muestra y diseñando un flujo de trabajo para cada objetivo. Se utilizan todas las técnicas y equipamiento disponibles, se implementan mejoras en los protocolos, y se interpretan y analizan los resultados obtenidos. La cartera de servicios (a grupos del CBM-SO y a externos) es amplia, desde la confirmación de la secuencia de una proteína recombinante, a la cuantificación relativa de proteomas, la caracterización de modificaciones post-traduccionales y de mutaciones, de interactomas o la secuenciación De Novo

Service description

Coomassie Gel Staining is a ready-to-use protein stain based on colloidal coomassie dye G-250 that provides nanogram-level detection and excellent clarity for polyacrylamide gels. This reagent stains only protein and, unlike traditional coomassie stains based on the R-250 form of the dye, allows bands to be viewed directly on the gel during the staining process. After staining, an optional water-wash step further enhances staining sensitivity and yields a clear background. Protocol: 1. Wash gel three times with methanol-acetic in water (50:10:40) 15 minutes. 2. Wash gel three times with MQ water 15 minutes. 3. Add colloidal coomassie 1 hour/over night. 4. For increased clarity use water wash enhancement step (1 hour). User instructions: 1. The solvents used will be high analytical grade. 2. The recipient should be preferably new. 3. Users should exercise extreme caution in handling to prevent contamination with keratins. 4. Users take the gel to the facility in methanol-acetic-water (50:10:40).

Options and prices chart
Options	Unit	Public Sector	Other customers
Coomassie Gel Staining	Gel	15.17 €	20.04 €

Service description

Conventional method for general and fast visualization of proteins separated by gel electrophoresis previous to their identification by MS. It is less sensitive than silver staining. Coomassie brilliant blue is an organic dye that forms strong, non-covalent complexes with proteins by combination of Van der Waals forces and its interaction with basic amino acids like arginine, lysine and histidine, as well as with tyrosine. The incorporation of the dye is proportional to the protein quantity. The affinity of the stain depends on the protein and correlates with its contents in basic amino acids. For staining, Coomassie brilliant blue is added to the gel dissolved in methanol/water/acetic acid in a 50/40/10 proportion, it has to be incubated for a period of 5-20 minutes and at the end the gel is destained with methanol/water/acetic acid in a 10/80/10 proportion for a period of 1-24 hours.This method allows to detect from 30 to 100ng of protein. Sensitivity of this conventional method can be improved by using high temperatures while staining and de-staining, or by using colloidal Coomassie brilliant blue. This last method allows protein detection in the range of 8-50ng, although it takes aprox. 1 to 2 days to be completed. For colloidad Coomassie staining, sample in the gel is submitted to a fixing step with methanol/water/acetic acid. Then the gel is incubated overnight with a Coomassie solution containing ammonium sulphate and phosphoric acid. Water is used to destain and the process can be accelerated by using temperatures that range from 45 to 55 grades.

Options and prices chart
Options	Unit	Public Sector	Other customers
Coomassie Gel Staining. Mini Gels.	Gel	31.43 €	34.42 €

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