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.
F2426
EZview™ Red ANTI-FLAG® M2 Affinity Gel
clone M2
Synonym(s):
Monoclonal ANTI-FLAG® M2 antibody produced in mouse, Anti-ddddk, Anti-dykddddk
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1 ML
$691.00
About This Item
UNSPSC Code:
12352203
NACRES:
NA.32
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clone
M2, monoclonal
analyte chemical class(es)
proteins
technique(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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This Item | A2220 | M8823 | F1804 |
|---|---|---|---|
| clone M2, monoclonal | clone M2, monoclonal | clone M2, monoclonal | clone M2, monoclonal |
| shipped in wet ice | shipped in wet ice | shipped in wet ice | shipped in wet ice |
| storage temp. −20°C | storage temp. −20°C | storage temp. −20°C | storage temp. −20°C |
| technique(s) affinity chromatography: suitable, immunoprecipitation (IP): suitable | technique(s) affinity chromatography: suitable, immunoprecipitation (IP): suitable | technique(s) affinity chromatography: suitable, immunoprecipitation (IP): suitable | technique(s) - |
| isotype IgG1 | isotype IgG1 | isotype IgG1 | isotype IgG1 |
| analyte chemical class(es) proteins | analyte chemical class(es) proteins | analyte chemical class(es) proteins | analyte chemical class(es) - |
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
Learn more product details in our FLAG® application portal.
Elution - FLAG peptide, Glycine, pH 3.5, 3x FLAG peptide
Learn more product details in our FLAG® application 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 Merck KGaA, Darmstadt, Germany
EZview is a trademark of Sigma-Aldrich Co. LLC
FLAG is a registered trademark of Merck KGaA, Darmstadt, Germany
Disclaimer
FLAG® Affinity Gels, FLAG® tag, 3xFLAG®tag, DYKDDDDK tag
Storage Class
10 - Combustible liquids
wgk_germany
WGK 3
flash_point_f
Not applicable
flash_point_c
Not applicable
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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?
1 answer-
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What is the binding capacity of the Product F2426, EZview™ Red ANTI-FLAG® M2 Affinity Gel clone M2, resin?
1 answer-
The binding capacity of the resin must be ? 0.6 mg/mL to meet specifications. This capacity will vary from lot to lot.
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When using Product F2426, EZview™ Red ANTI-FLAG® M2 Affinity Gel clone M2, how can I elute my protein?
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.
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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?
1 answer-
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.
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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?
1 answer-
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.
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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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