Anti-Flag M1 antibody is specific for a calcium complex of the FLAG peptide. M1 and M2 are different clones. The ANTI-FLAG M1 monoclonal antibody binds to proteins with a FLAG marker at the free N-terminus. The ANTI-FLAG M2 monoclonal antibody recognizes the FLAG peptide sequence at the N-terminus, Met-N-terminus,C-terminus, and internal sites of the fusion protein.
A4596
ANTI-FLAG® M1 Agarose Affinity Gel
Synonym(s):
Anti-ddddk, Anti-dykddddk
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About This Item
NACRES:
NA.32
UNSPSC Code:
12352203
Conjugate:
agarose conjugate
Species reactivity:
-
Application:
—
Citations:
81
Pricing and availability is not currently available.
conjugate
agarose conjugate
Quality Segment
antibody product type
primary antibodies
form
suspension
isotype
IgG12b
capacity
≥0.6 mg/mL, gel binding capacity
storage temp.
−20°C
General description
ANTI-FLAG® M1 Agarose Affinity Gel is a purified IgG2B monoclonal antibody covalently attached to agarose.
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
Learn more product details in our FLAG® application portal.
Affinity gel is for calcium mediated purification of N-terminal FLAG fusion proteins.
immunoprecipitation (IP): suitable
Elution - FLAG peptide, Glycine, pH 3.5 EDTA
Learn more product details in our FLAG® application portal.
Biochem/physiol Actions
Binding specificity: Free N-Terminus of FLAG sequence
N-Asp-Tyr-Lys-Asp-Asp-Asp-Asp-Lys-C
N-Asp-Tyr-Lys-Asp-Asp-Asp-Asp-Lys-C
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.
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
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.
Legal Information
ANTI-FLAG is a registered trademark of Merck KGaA, Darmstadt, Germany
FLAG is a registered trademark of Merck KGaA, Darmstadt, Germany
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This Item | |||
|---|---|---|---|
| conjugate agarose conjugate | conjugate agarose conjugate | conjugate - | conjugate unconjugated |
| storage temp. −20°C | storage temp. 2-8°C | storage temp. −20°C | storage temp. −20°C |
| form suspension | form - | form - | form buffered aqueous solution (10 mM sodium phosphate, 150 mM NaCl, pH 7.4, containing 0.02% sodium azide) |
| isotype IgG12b | isotype - | isotype IgG1 | isotype IgG1 |
| Quality Level 200 | Quality Level 200 | Quality Level 200 | Quality Level 200 |
| capacity ≥0.6 mg/mL, gel binding capacity | capacity 2.5 nmol/mL, gel (V5 fusion protein) | capacity ≥0.6 mg/mL, gel binding capacity | capacity - |
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Storage Class
10 - Combustible liquids
wgk
WGK 3
flash_point_f
Not applicable
flash_point_c
Not applicable
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Bonjour, pouvez vous m'expliquer pour quoi la résine anti Flag M1 est calcium dépendante contrairement à la résine antiFlag M2 sachant que les séquences de reconnaissances sont les mêmes?
1 answer-
Helpful?
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How much calcium is required for binding of my FLAG-tagged protein to Product A4596, ANTI-FLAG® M1 Agarose Affinity Gel?
1 answer-
10 mM calcium chloride is recommended for binding.
Helpful?
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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.
Helpful?
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What reagents are compatible with Product A4596, ANTI-FLAG® M1 Agarose Affinity Gel?
1 answer-
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.
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Can I use Phosphate Buffered Saline (PBS) instead of Tris Buffered Saline (TBS) with Product A4596, ANTI-FLAG® M1 Agarose Affinity Gel?
1 answer-
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.
Helpful?
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Will Product A4596, ANTI-FLAG® M1 Agarose Affinity Gel, work for my C-terminal FLAG-tagged protein?
1 answer-
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.
Helpful?
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