F2426

Millipore

EZview Red ANTI-FLAG® M2 Affinity Gel

clone M2

Synonym(s):
Anti-ddddk, Anti-dykddddk, Monoclonal ANTI-FLAG M2 antibody produced in mouse
NACRES:
NA.32

Quality Level

clone

M2, monoclonal

analyte chemical class(es)

proteins (FLAG® Affinity Gels, FLAG® tag, 3xFLAG® tag, DYKDDDDK tag)

application(s)

affinity chromatography: suitable
immunoprecipitation (IP): suitable

matrix

4% agarose bead; 45-165μm bead size

isotype

IgG1

capacity

≥0.6 mg/mL, gel binding capacity

shipped in

wet ice

storage temp.

−20°C

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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.

Specificity

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

Application

Immunoprecipitation (IP) of FLAG- and 3xFLAG-tagged fusion proteins.

Elution - FLAG peptide, Glycine, pH 3.5, 3x FLAG peptide

Browse additional application references in our
FLAG® Literature portal.

Physical form

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

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

RIDADR

NONH for all modes of transport

WGK Germany

WGK 3

Flash Point(F)

Not applicable

Flash Point(C)

Not applicable

  1. 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.

  2. 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.

  3. 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.

  4. 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.

  5. 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.

  6. 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. 

  7. 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.

  8. 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.

  9. 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. 

  10. My question is not addressed here, how can I contact Technical Service for assistance?

    Ask a Scientist here.

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Protocols
Protocol for immunoprecipitation (IP) of FLAG fusion proteins using M2 monoclonal antibody 4% agarose affinity gels
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