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F2426 Sigma

EZview Red ANTI-FLAG® M2 Affinity Gel

clone M2

Synonym: Anti-ddddk, Anti-dykddddk, Monoclonal ANTI-FLAG® M2 antibody produced in mouse

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Description

Frequently Asked Questions

Frequently Asked Questions are available for this Product.

Application

Immunoprecipitation of FLAG- and 3xFLAG-tagged fusion proteins.

Browse additional application references in our FLAG® Literature portal.

Features and Benefits

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Evaluate our antibodies with complete peace of mind. If the antibody does not perform in your application, we will issue a full credit or replacement antibody. Learn more.

Specificity

Suitable for purifying N-terminal, Met-N-terminal, C-terminal FLAG fusion proteins, 3xFLAG fusion proteins.

Physical form

1:1 (v/v) suspension in PBS containing 50% glycerol and 15 ppm Kathon

General description

EZ view Red Anti-FLAG M2 Affinity Gel is a resin that consists of Anti-FLAG M2 antibody, covalently bonded to 4% Agarose beads. The affinity gel is used to bind FLAG fusion proteins to samples, such as cell lysates and tissue, for purification of FLAG-tagged proteins in preparation of immunoprecipitation assays. Red dye enhances visability for more efficient results. Agarose beads bind at N-terminal, Met-N-terminal and C-terminal FLAG fusion proteins, 3xFLAG-tagged fusion proteins.

Legal Information

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

EZview is a trademark of Sigma-Aldrich Co. LLC

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

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Safety & Documentation

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.
When using Product F2426, EZview™ Red ANTI-FLAG® M2 Affinity Gel clone M2, should I use a 3X FLAG peptide or a 1X FLAG peptide to elute my protein?
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.  To view this information in the F2426 Technical Bulletin, click on the following link:  http://www.sigmaaldrich.com/etc/medialib/docs/Sigma/Bulletin/f2426bul.Par.0001.File.tmp/f2426bul.pdf
When using Product F2426, EZview™ Red ANTI-FLAG® M2 Affinity Gel clone M2, how can I elute my protein?
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.  To view this information in the F2426 Technical Bulletin, click on the following link:  http://www.sigmaaldrich.com/etc/medialib/docs/Sigma/Bulletin/f2426bul.Par.0001.File.tmp/f2426bul.pdf
When using Product F2426, EZview™ Red ANTI-FLAG® M2 Affinity Gel clone M2, I see bands at 20-25 kDa and 50-60 kDa appearing in my Westerns that are not my FLAG-tagged protein. How can I prevent this?
As a result of the conjugation, there may be some M2 antibody that is not conjugated to the resin, but is associated with the resin and may appear in acid elutions as heavy and light chain when using the anti-mouse IgG conjugated secondary antibody.  We recommend a acid wash (0.1 M glycine-HCL pH 3.5) and neutralization of the resin (do not allow the acid wash to sit on the resin longer than 20 minutes) prior to applying the lysate.  Another way to avoid this is to use a directly conjugated FLAG antibody for detection such as product A8592 ant-FLAG M2 HRP, or the rabbit anti-FLAG polyclonal antibody, F7425.  To view this information in the F2426 Technical Bulletin, click on the following link:  http://www.sigmaaldrich.com/etc/medialib/docs/Sigma/Bulletin/f2426bul.Par.0001.File.tmp/f2426bul.pdf
When using Product F2426, EZview™ Red ANTI-FLAG® M2 Affinity Gel clone M2, I have a lot of non-specific proteins that are eluting with my FLAG-tagged protein. How can I get rid of these?
One way to remove non-specific proteins is to pre-bind the protein lysate with unconjugated resin.  We recommend product 4B200 for this purpose. Other methods 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.  To view this information in the F2426 Technical Bulletin, click on the following link:  http://www.sigmaaldrich.com/etc/medialib/docs/Sigma/Bulletin/f2426bul.Par.0001.File.tmp/f2426bul.pdf
What is the binding capacity of the Product F2426, EZview™ Red ANTI-FLAG® M2 Affinity Gel clone M2, resin?
The binding capacity of the resin must be   ≥ 0.6 mg/mL to meet specifications.  This capacity will vary from lot to lot.  To view this information in the Technical Bulletin, click on the following link:   http://www.sigmaaldrich.com/etc/medialib/docs/Sigma/Bulletin/f2426bul.Par.0001.File.tmp/f2426bul.pdf
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
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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