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A4596 Millipore

ANTI-FLAG® M1 Agarose Affinity Gel

Synonym: Anti-ddddk, Anti-dykddddk

  •  NACRES NA.32

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Properties

Related Categories Alphabetical Index, Antibodies, Antibodies to Peptide and Protein Fusion Tags, FK-FO, FLAG Affinity Gels,
conjugate   agarose conjugate
storage temp.   −20°C
isotype   IgG12b
Quality Level   200
antibody product type   primary antibodies
capacity   ≥0.6 mg/mL, gel binding capacity

Description

General description

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

Specificity

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

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.

immunoprecipitation (IP): suitable

Elution - FLAG peptide, Glycine, pH 3.5 EDTA

Browse additional application references in our FLAG® Literature portal.

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

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.

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.

Safety & Documentation

Safety Information

RIDADR 
NONH for all modes of transport
WGK Germany 
WGK 3
Flash Point(F) 
Not applicable
Flash Point(C) 
Not applicable

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.
How do I get lot-specific information or a Certificate of Analysis?
The lot specific COA document can be found by entering the lot number above under the "Documents" section.
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.
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.
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.
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.
My question is not addressed here, how can I contact Technical Service for assistance?
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Protocols & Articles

Articles

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Peer-Reviewed Papers
15

References

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