Detailed equilibration and regeneration protocols for this product are provided in the Product Information Sheet:
http://sigmaaldrich.com/deepweb/assets/sigmaaldrich/product/documents/288/183/a2220bul-ms.pdf
A2220
Gel ANTI-FLAG® M2 Affinity
ANTI-FLAG® M2 Affinity Gel
purified immunoglobulin, buffered aqueous glycerol solution
Sinónimos:
ANTI-FLAG® M2 monoclonal antibody produced in mouse, Gel de agarosa ANTI-FLAG® M2 Affinity, Anti-ddddk, Anti-dykddddk
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1 ML
MXP 11,664.00
5 ML
MXP 36,449.00
10 ML
MXP 54,423.00
25 ML
MXP 114,892.00
2 X 25 ML
MXP 179,393.00
4 X 25 ML
MXP 346,673.00
Seleccione un Tamaño
Cambiar Vistas
1 ML
MXP 11,664.00
5 ML
MXP 36,449.00
10 ML
MXP 54,423.00
25 ML
MXP 114,892.00
2 X 25 ML
MXP 179,393.00
4 X 25 ML
MXP 346,673.00
About This Item
Código UNSPSC:
12352203
NACRES:
NA.32
Productos recomendados
conjugado
agarose conjugate
Nivel de calidad
forma del anticuerpo
purified immunoglobulin
tipo de anticuerpo
primary antibodies
clon
M2, monoclonal
Formulario
buffered aqueous glycerol solution
clases químicas de analitos
proteins
técnicas
affinity chromatography: suitable
immunoprecipitation (IP): suitable
Matriz
(4% agarose bead; 45-165μm bead size)
isotipo
IgG1
capacidad
>0.6 mg/mL, resin binding capacity (FLAG-BAP)
Condiciones de envío
wet ice
temp. de almacenamiento
−20°C
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Descripción general
M2 anti-FLAG purificado en gel de afinidad, es un anticuerpo monoclonal que está covalentemente unido a la agarosa. El anticuerpo se une a FLAG en los extremos N-terminal, Met-N-terminal, C-terminal y ubicaciones internas de las proteínas de fusión. La unión no es dependiente del calcio.
Elución - Péptido FLAG®, glicina, pH 3,5, 3x péptido FLAG®
Elución - Péptido FLAG®, glicina, pH 3,5, 3x péptido FLAG®
Péptido FLAG®, glicina, pH 3,5, péptido 3x FLAG®
Inmunógeno
DYKDDDDK
Aplicación
El gel de afinidad anti-FLAG® M2 se ha utilizado para inmunoelectrotransferencia (western blotting) e inmunoprecipitación, y para la purificación de las proteínas de fusión FLAG.[1][2][3]
Conozca más detalles sobre el producto en nuestro portal de la aplicación FLAG® .
Conozca más detalles sobre el producto en nuestro portal de la aplicación FLAG® .
Forma física
Suspensión en una disolución salina tamponada que contenga acida como conservante y glicerol al 50 %
Información legal
ANTI-FLAG is a registered trademark of Merck KGaA, Darmstadt, Germany
FLAG is a registered trademark of Merck KGaA, Darmstadt, Germany
Cláusula de descargo de responsabilidad
Etiqueta FLAG®, 3x FLAG®, etiqueta DYKDDDDK
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Referencia del producto
Descripción
Precios
Código de clase de almacenamiento
10 - Combustible liquids
Clase de riesgo para el agua (WGK)
WGK 1
Punto de inflamabilidad (°F)
Not applicable
Punto de inflamabilidad (°C)
Not applicable
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Certificados de análisis (COA)
Lot/Batch Number
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Artículos
The FLAG® Expression System is a proven method to express, purify and detect recombinant fusion proteins. Sigma®, the proven provider of FLAG®, now offers a magnetic bead for immunoprecipitation, protein purification, and the study of protein-protein interactions. The ANTI-FLAG® M2 Magnetic Bead is composed of murine derived, anti-FLAG® M2 monoclonal antibody attached to superparamagnetic iron impregated 4% agarose beads, with an average diameter of 50 µm. The M2 antibody is capable of binding to fusion proteins containing a FLAG peptide sequence at the N-terminus, Met-N-terminus, or C-terminus locations in mammalian, bacterial, and plant extracts.
Protocolos
Protocol for immunoprecipitation (IP) of FLAG fusion proteins using M2 monoclonal antibody 4% agarose affinity gels
Contenido relacionado
Técnicas, reactivos y protocolos de purificación de proteínas para purificar proteínas recombinantes utilizando métodos como la cromatografía de intercambio iónico, de exclusión por tamaño y de afinidad.
Protein purification techniques, reagents, and protocols for purifying recombinant proteins using methods including, ion-exchange, size-exclusion, and protein affinity chromatography.
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How do I equilibrate the resin and how can I clean it for re-use?
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May I ask a question about the product? Does "1mL" mean 1 mL Slurry plus 50% glycerol, which is 2 mL in total? Same as other (e.g., 10 mL = 50% slurry with total 20 mL volume)
1 answer-
The "1mL" indicates that this product is supplied at a volume of 1 milliliter. This ANTI-FLAG M2 affinity gel is supplied as a 50% suspension in 50% glycerol with 10 mM sodium phosphate and 150 mM sodium chloride, pH 7.4, containing 0.02% (w/v) sodium azide (PBS/A).
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What is the bead size for this product?
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As stated in the 'PROPERTIES' section of the product page, the bead size range is 45-165 μm.
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Is there a generic agarose bead available for purchase that can be used as a control to test for non-specific binding, in the case that something is binding to the agarose bead itself rather than the tag? If it is not listed on the site, is there a possibility of obtaining generic agarose beads through alternative means? Alternatively, could any information be provided to determine if another provider's generic agarose beads are produced in a similar manner for this specific purpose?
1 answer-
Yes, there are two options available for a generic agarose bead as a control to test for non-specific binding. One option is Product Number. 4B200, Sepharose® 4B, which can be used for sample preclearing and is equivalent to the beads in the FLAG affinity product. The other option is Product Number: A0919, Mouse IgG−Agarose, which can serve as a conjugated antibody bead control.
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Is there a protocol for regeneration?
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Please see the link below to the product data-sheet. Instructions state that the column should be regenerated immediately after use by washing with three column volumes of 0.1 M glycine HCI, pH 3.5. The column should be immediately re-equilibrated in TBS until the effluent is at neutral pH:
https://www.sigmaaldrich.com/deepweb/assets/sigmaaldrich/product/documents/288/183/a2220bul-ms.pdf
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How should I elute my protein when using Product No. A2220, ANTI-FLAG® M2 Affinity Gel?
1 answer-
Elution with the peptide is the most gentle method. Acid elution (0.1 M glycine-HCL pH 3.5) is a more stringent method of elution, and should be evaluated for its effect on your protein if it is to be used in downstream applications. Boiling the resin in sample buffer is the most denaturing condition. If this condition is used, the resin cannot be re-used, due to the presence of SDS and/or reducing agents. The elution information can be viewed on A2220 product information sheet (under Documents, above).
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When using ANTI-FLAG M2 Affinity Gel, Product A2220, should I use a 3X FLAG peptide or a 1X FLAG peptide to elute my protein?
1 answer-
If you have a 3X FLAG-tagged protein, then you will need to use the 3X FLAG peptide. If you have a 1X FLAG-tagged protein, you can use the 1X FLAG peptide or the 3X FLAG peptide. We have not noticed a significant difference in elution efficiency by using a 3X FLAG peptide on a 1X FLAG-tagged protein.
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What is the Department of Transportation shipping information for this product?
1 answer-
Transportation information can be found in Section 14 of the product's (M)SDS.To access the shipping information for this material, use the link on the product detail page for the product.
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Is my lysis buffer compatible with Product No. A2220, ANTI-FLAG® M2 Affinity Gel?
1 answer-
The A2220 product information sheet (under Documents, above) contains a reagent compatibility table located on page 6. We do not recommend addition of SDS or reducing agents such as DTT, DTE or 2-mercaptoethanol to the resin. If you have a detergent listed in the table in a higher than recommended concentration, we recommend trying to dilute the sample before applying to the resin.
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I am using Product No. A2220, ANTI-FLAG® M2 Affinity Gel, and have a lot of non-specific proteins that are eluting with my FLAG®-tagged protein. How can I get rid of these?
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The product bulletin for Product A2220, ANTI-FLAG® M2 affinity gel indicates:Pre-clear lysate with Mouse IgG-Agarose (Product A0919) to remove nonspecific binding proteins. Alternatively, you can use the unconjugated resin (Product 4B200) for this purpose. Other methods to remove non-specific binding from the resin would be to increase the stringency of the washes by increasing salt concentration (the resin can tolerate up to 1M NaCl) or including detergents that are compatible with the resin.
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