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A4596 Sigma

ANTI-FLAG® M1 Agarose Affinity Gel

Synonym: Anti-ddddk, Anti-dykddddk

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Properties

Related Categories Alphabetical Index, Antibodies, Antibodies to Peptide and Protein Fusion Tags, FK-FO, FLAG Affinity Gels,
isotype   IgG12b
capacity   ≥0.6 mg/mL, gel binding capacity
conjugate   agarose conjugate
storage temp.   −20°C

Description

Frequently Asked Questions

Frequently Asked Questions are available for this Product.

Application

For purification of N-terminal FLAG fusion proteins. Since binding is Ca2+-dependent, proteins can be eluted with a buffer containing EDTA, as well as by the standard methods using either FLAG peptide or glycine-HCl buffer, pH 3. ANTI-FLAG M1 does not bind to Met-FLAG fusion proteins, so this resin is not appropriate for purifying unprocessed, cytoplasmically expressed fusion proteins.

Affinity gel is for calcium mediated purification of N-terminal FLAG fusion proteins.

Browse additional application references in our FLAG® Literature portal.

Features and Benefits

• Typically purify fusion proteins from crude lysates to single band purity in just one chromatography step.
• Fusion protein may be eluted from affinity resin by mild elution with EDTA.
• A solution of FLAG peptide can be used for gentle, non-denaturing elution of FLAG fusion proteins.

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General description

ANTI-FLAG® M1 Agarose Affinity Gel is a purified mouse IgG2B monoclonal antibody covalently attached to agarose.

Other Notes

ANTI-FLAG® M1 does not bind to Met-FLAG fusion proteins, so this resin is not appropriate for purifying unprocessed, cytoplasmically expressed fusion proteins.

Specificity

Binding specificity: Free N-Terminus of FLAG sequence
N-Asp-Tyr-Lys-Asp-Asp-Asp-Asp-Lys-C

Physical form

Suspension of beaded agarose in 50% glycerol containing 10 mM sodium phosphate, 150 mM NaCl, pH 7.4, 0.02% (w/v) sodium azide

Legal Information

ANTI-FLAG is a registered trademark of Sigma-Aldrich Co. LLC

FLAG is a registered trademark of Sigma-Aldrich Co. LLC

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Safety Information

RIDADR 
NONH for all modes of transport
WGK Germany 
3

Frequently Asked Questions

Which document(s) contains shelf-life or expiration date information for a given product?
If available for a given product, the recommended re-test date or the expiration date can be found on the Certificate of Analysis. These documents are located on the product detail page under Useful Links & Tools. Click on the following link to search for a Certificate of Analysis. Please click the following link to see the details on our Product Dating Information.
How do I get lot-specific information or a Certificate of Analysis?
A Certificate of Analysis is available by lot number and can be obtained through our Advanced Search Option: http://www.sigmaaldrich.com/catalog/AdvancedSearchPage.do
Will Product A4596, ANTI-FLAG® M1 Agarose Affinity Gel, work for my C-terminal FLAG-tagged protein?
No.  The M1 FLAG antibody resin will only bind proteins that are free N-terminal FLAG tagged proteins.  It will not recognize C-terminal FLAG tagged proteins or Met-N-terminal FLAG tagged proteins.  For more information on the M1 antibody see the following link:  http://www.sigmaaldrich.com/sigma/datasheet/a4596dat.pdf
Can I use Phosphate Buffered Saline (PBS) instead of Tris Buffered Saline (TBS) with Product A4596, ANTI-FLAG® M1 Agarose Affinity Gel?
We do not recommend the use of phosphate buffers since calcium is required for binding.  This can result in the formation of insoluble calcium phosphates in the presence of PBS.
How much calcium is required for binding of my FLAG-tagged protein to Product A4596, ANTI-FLAG® M1 Agarose Affinity Gel?
10 mM calcium chloride is recommended for binding.  For more information please see the product datasheet at this link:  http://www.sigmaaldrich.com/sigma/datasheet/a4596dat.pdf
What reagents are compatible with Product A4596, ANTI-FLAG® M1 Agarose Affinity Gel?
The ANTI-FLAG M1 Affinity Gel is resistant to the following detergents: 5.0 % Tween-20; 5.0 % Triton X-100; 0.1 % NP-40; 0.1 % CHAPS and 0.2 % digitonin. It can also be used with 1.0 M NaCl or 1.0 M urea. Do not use the gel in the presence of SDS, deoxycholate or guanidine HCl.  This information can be found in the product datasheet at the following link: http://www.sigmaaldrich.com/etc/medialib/docs/Sigma/datasheet6/a4596dat.Par.0001.File.tmp/a4596dat.pdf
How do I find price and availability?
There are several ways to find pricing and availability for our products.  Once you log onto our website, you will find the price and availability displayed on the product detail page. You can contact any of our Customer Sales and Service offices to receive a quote.  USA customers:  1-800-325-3010 or view local office numbers. 
What is the Department of Transportation shipping information for this product?
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, or search here. 
My question is not addressed here, how can I contact Technical Service for assistance?
Use the option to the right to "Ask a Question" by email of a Technical Service Scientist.
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Protocols & Articles

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