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Insist on the Original & Proven Performer...
The FLAG Expression System is a proven method to express, purify and detect recombinant
fusion proteins. FLAG and 3xFLAG are useful in Western blotting, immunocytochemistry, immunoprecipitation, flow cytometry, protein purification, and in the study of protein-protein interactions, cell ultrastructure, and protein localization. These small hydrophilic tags significantly improve detection and purification of recombinant fusion proteins when used with our highly specific and sensitive ANTI-FLAG® antibodies.
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| FLAG Vectors |
|
| Bacterial Expression Vectors |
|
| Product # |
Product Name |
Package Size |
| E5769 |
pFLAG-ATS™ |
10 µg |
| E5644 |
pFLAG-MAC™ |
10 µg |
| E5269 |
pFLAG-CTS™ |
10 µg |
| E5394 |
pFLAG-CTC™ |
10 µg |
| E7898 |
pFLAG-Shift12c™ |
10 µg |
| P1118 |
pT7-FLAG™-1 |
10 µg |
| P1243 |
pT7-FLAG™-2 |
10 µg |
| E5280 |
pT7-FLAG™-MAT-1 |
10 µg |
| E4905 |
pT7-MAT-FLAG™-2 |
10 µg |
| E5155 |
pT7-MAT-FLAG™-1 |
10 µg |
| E5030 |
pT7-FLAG-MAT™-2 |
10 µg |
| P9618 |
pT7-FLAG™-3 |
10 µg |
| P9743 |
pT7-FLAG™-4 |
10 µg |
| Mammalian and BICEP™ Expression Vectors |
|
| Product # |
Product Name |
Package Size |
| E7398 |
pFLAG-CMV™-2 |
20 µg |
| E7901 |
pFLAG-CMV™-5.1 |
20 µg |
| E3762 |
pFLAG-CMV™-5a,b,c |
1 set |
| E2275 |
pFLAG-CMV™-6a,b,c |
1 set |
| E4026 |
p3xFLAG-CMV™-7.1 |
20 µg |
| E4151 |
p3xFLAG-CMV™-8 |
20 µg |
| E5526 |
pFLAG-myc-CMV™-19 |
20 µg |
| E5651 |
pFLAG-myc-CMV™-20 |
20 µg |
| E6026 |
p3xFLAG-myc-CMV™-23 |
20 µg |
| E6151 |
p3xFLAG-myc-CMV™-24 |
20 µg |
| C5864 |
pCMV-FLAG-MAT™-1 |
20 µg |
| C5989 |
pCMV-MAT-FLAG™-1 |
20 µg |
| C6114 |
pCMV-FLAG-MAT™-2 |
20 µg |
| E7029 |
pBICEP-CMV™-3 |
20 µg |
| E5905 |
pCMV-BICEP™-4 |
20 µg |
| Stable Expression |
|
| Product # |
Product Name |
Package Size |
| E8770 |
pFLAG-CMV™-3 |
20 µg |
| E1775 |
pFLAG-CMV™-4 |
20 µg |
| E4276 |
p3xFLAG-CMV™-9 |
20 µg |
| E4401 |
p3xFLAG-CMV™-10 |
20 µg |
| E4776 |
p3xFLAG-CMV™-13 |
20 µg |
| E4901 |
p3xFLAG-CMV™-14 |
20 µg |
| E5776 |
pFLAG-myc-CMV™-21 |
20 µg |
| E5901 |
pFLAG-myc-CMV™-22 |
20 µg |
| E6276 |
p3xFLAG-myc-CMV™-25 |
20 µg |
| E6401 |
p3xFLAG-myc-CMV™-26 |
20 µg |
| E0779 |
pBICEP-CMV™-1 |
20 µg |
| E0904 |
pBICEP-CMV™-2 |
20 µg |
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| FLAG Affinity Gels |
|
| Product # |
Package Size |
Product Name |
Characteristics |
Applications |
| A4596 |
1 ml
5 ml
10 ml
25 ml |
ANTI-FLAG® M1 Agarose Affinity Gel |
Specificity: N-terminal FLAG fusion proteins. Binding Ca2+-dependent; the complex dissociates in the absence of calcium ions. Does not bind to Met-FLAG fusion proteins, will not recognize unprocessed, cytoplasmically expressed proteins.
Binding Capacity: > 0.6 mg protein per ml gel, binding requires Ca2+
Elution: FLAG peptide; glycine, pH 3.5; EDTA
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 |
• Immunoprecipitation
• Purification of N-terminal FLAG fusion proteins |
| A2220 |
1 ml
5 ml
10 ml
25 ml |
ANTI-FLAG® M2 Agarose Affinity Gel (Freezer Safe) |
Specificity: N-terminal, Met-N-terminal, C-terminal FLAG fusion proteins, 3xFLAG fusion proteins
Binding Capacity: > 0.6 mg per ml gel
Elution: FLAG peptide; glycine, pH 3.5; 3xFLAG Peptide
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. |
• Immunoprecipitation
• Purification of FLAG and 3xFLAG fusion proteins |
| F2426 |
1 ml
5 x 1 ml |
EZview™ Red ANTI-FLAG® M2 Affinity Gel |
Specificity: N-terminal, Met-N-terminal, C-terminal FLAG fusion proteins, 3xFLAG fusion proteins
Binding Capacity: > 0.6 mg per ml gel
Elution: FLAG peptide glycine, pH 3.5; 3xFLAG peptide
Form: Suspension of red colored beaded agarose in phosphate buffered saline containing 50% glycerol and 0.0015% Kathon® CG/IPCII as an antimicrobial preservative |
• Immunoprecipitation |
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| FLAG Peptides |
|
| Product # |
Package Size |
Product Name |
Characteristics |
Applications |
| F3290 |
4 mg
25 mg |
FLAG® Peptide |
Sequence: Asp-Tyr-Lys-Asp-Asp-Asp-Asp-Lys
MW: 1013
Form: Lyophilized powder |
• Elution of FLAG fusion proteins from the ANTI-FLAG M1 and M2 affinity resins
Working Concentration:
• 100 µg/ml is commonly used to elute FLAG fusion proteins |
| F4799 |
4 mg
25 mg |
3xFLAG® Peptide |
Sequence: Met-Asp-Tyr-Lys-Asp-His-
Asp-Gly-Asp-Tyr-Lys-Asp-His-Asp-Ile-Asp-Tyr-Lys-
Asp-Asp-Asp-Asp-Lys
Note: The Asp-Tyr-Lys-Xaa-Xaa-Asp motif is repeated three times in the peptide; the eight amino acids at the C-terminus make up the classic FLAG sequence (Asp-Tyr-Lys-Asp-Asp-Asp-Asp-Lys).
MW: 2861.9
Form: Lyophilized powder |
• Elution of 3xFLAG fusion proteins from ANTI-FLAG M2 affinity gels
Working Concentration:
• 100 µg/ml is commonly used to elute 3xFLAG fusion proteins |
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| FLAG Antibodies |
|
| Product # |
Package Size |
Product Name |
Characteristics |
Applications |
| F1804 |
200 µg
1 mg
5 mg |
New!
ANTI-FLAG® M2 Monoclonal Antibody, Affinity Purified IgG |
Specificity: N-terminal FLAG, Met-N-terminal, Carboxy-terminal, or internal. Binding is not Ca2+- dependent.
Form: Solution in 50% glycerol, 10 mM sodium phosphate, 150 mM NaCl, pH 7.4. The formulation contains no antimicrobial preservatives. |
• Immunoprecipitation
• Immunocytochemistry
• Western blotting
• EIA
Working Dilution:
• 1 µg/ml by indirect Western blotting (chemiluminescent) |
| F3040 |
200 µg
1 mg
5 mg |
ANTI-FLAG® M1 Monoclonal Antibody, Purified IgG |
Specificity: N-terminal FLAG. Binding is Ca2+- dependent; the complex dissociates in the absence of calcium ions. Does not bind to Met-FLAG fusion proteins; will not recognize unprocessed, cytoplasmically expressed proteins.
Form: Solution in 10 mM sodium phosphate, 150 mM NaCl, pH 7.4, containing 0.02% sodium azide. |
• Immunoprecipitation
• Immunocytochemistry
• Western blotting
• EIA
Working Dilution:
• 10 µg/ml by indirect Western blotting (chemiluminescent) |
| F3165 |
200 µg
1 mg
5 mg |
ANTI-FLAG® M2 Monoclonal Antibody, Purified IgG |
Specificity: N-terminal, Met-N-terminal, Carboxy-terminal, or internal. Binding is not Ca2+-dependent.
Form: Solution in 10 mM sodium phosphate, 150 mM NaCl, pH 7.4, containing 0.02% sodium azide. |
• Immunoprecipitation
• Immunocytochemistry
• Western blotting
• EIA
Working Dilution:
• 10 µg/ml by indirect Western blotting (chemiluminescent) |
| F4042 |
200 µg
1 mg
5 mg |
ANTI-FLAG® M5 Monoclonal Antibody |
Specificity: Met-N-terminal FLAG. Useful for detecting cytoplasmically expressed Met-FLAG fusion proteins in mammalian crude cell extracts, but not recommended for fusion proteins expressed in E. coli. Binding is not Ca2+-dependent.
Form: Solution in 10 mM sodium phosphate, 150 mM NaCl, pH 7.4, containing 0.02% sodium azide. |
• Western blotting
Working Dilution:
• 10 µg/ml by indirect Western blotting (chemiluminescent) |
| F2555 |
200 µl |
Monoclonal ANTI-FLAG® Antibody produced in rabbit, ascites fluid |
Specificity: N-terminal and C-terminal FLAG fusion proteins.
Form: Provided as ascites fluid containing 15 mM sodium azide as a preservative. |
• Immunocytochemistry
• Western blotting
• EIA
Working Dilution:
1:1000 - 1:2000 by indirect Western blotting (chemiluminescent) |
| F7425 |
200 µg |
Rabbit ANTI-FLAG® Polyclonal Antibody |
Specificity: Reacts with N-terminal, Met-N-terminal, and C-terminal FLAG fusion proteins. Binding is not Ca2+-dependent.
Form: Solution of affinity isolated antibody in 10 mM phosphate buffered saline, pH 7.4, containing 1% bovine serum albumin and 15 mM sodium azide. |
• Immunoprecipitation
• Immunocytochemistry
• Western blotting
• Dot blotting
Working Dilution:
• 5 µg/ml by indirect immunofluorescence
• 2.5 µg/ml by indirect Western blotting (chemiluminescent) |
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| FLAG Antibody Conjugates |
|
| Product # |
Package Size |
Product Name |
Characteristics |
Applications |
| F9291 |
200 µg
1 mg
5 x 1 mg |
ANTI-FLAG® BioM2 Antibody, Biotin Conjugate |
Specificity: N-terminal, Met-N-terminal or C-terminal of FLAG fusion proteins. Binding is not Ca2+-dependent.
Form: Solution in 50% glycerol, 10 mM sodium phosphate, 150 mM NaCl, pH 7.4, containing 0.02% sodium azide. The conjugate protein concentration is approximately 1 mg/ml. |
• Western blotting
• Immunocytochemistry
Working Dilution:
• 10 µg/ml by indirect Western blotting (chemiluminescent) |
| F2922 |
200 µg
1 mg |
ANTI-FLAG® BioM5 Monoclonal Antibody, Biotin Conjugate |
Specificity: Met-N-terminal FLAG fusion proteins. Binding is not Ca2+-dependent. ANTI-FLAG BioM5 is not recommended for detection of FLAG fusion proteins in E. coli.
Form: Solution in 10 mM sodium phosphate, 150 mM NaCl, pH 7.4, containing 0.02% sodium azide. |
• Western blotting
• Immunocytochemistry
Working Dilution:
• 2 µg/ml by indirect Western blotting (chemiluminescent) |
| A9469 |
200 µg
1 mg
5 x 1 mg |
ANTI-FLAG® M2-Alkaline Phosphatase |
Specificity: N-terminal, Met-N-terminal, or C-terminal of FLAG fusion proteins. Especially useful in detection of FLAG fusion proteins expressed in murine host, where secondary anti-mouse antibodies may cause cross-reactivity. Binding is not Ca2+-dependent.
Form: Purified immunoglobulin solution in Tris buffered saline containing 50% glycerol plus stabilizer and preservative. The conjugate protein concentration is approximately 1 mg/ml. |
• Western blotting
• ELISA
Working Dilution:
• 1:20,000 by indirect ELISA |
| A8592 |
200 µg
1 mg
5 x 1 mg |
ANTI-FLAG® M2-Peroxidase |
Specificity: N-terminal, Met-N-terminal, or C-terminal of FLAG fusion proteins. Especially useful in detection of FLAG fusion proteins expressed in murine host, where secondary anti-mouse antibodies may cause cross-reactivity. Binding is not Ca2+-dependent.
Form: Solution in phosphate buffered saline containing 50% glycerol plus preservative and stabilizer. The conjugate protein concentration is approximately 1 mg/ml. |
• Immunocytochemistry
• Immunohistochemistry
• ELISA
• Western blotting
Working Dilution:
• 1:20,000 by indirect ELISA
• 1:100-1:1,000 by immunocytochemistry |
| F4049 |
200 µg
1 mg
5 x 1 mg |
ANTI-FLAG® M2 Monoclonal Antibody-FITC |
Specificity: N-terminal, Met-N-terminal, or C-terminal of FLAG fusion proteins. Binding is not Ca2+-dependent.
Form: Solution in 10 mM sodium phosphate, 150 mM NaCl, 1% bovine serum albumin, 0.1% sodium azide, pH 7.4. |
• Fluorescent immunocytochemistry
• Fluorescent immunohistochemistry
• Flow cytometry
Working Dilution:
• 10 µg/ml by indirect immunofluorescence using mammalian cells fixed with methanol:acetone |
| A9594 |
200 µg
1 mg
5 x 1 mg |
ANTI-FLAG® M2-Cy3™ Conjugate |
Specificity: N-terminal, Met-N-terminal, or C-terminal of FLAG fusion proteins. Especially useful in detection of FLAG fusion proteins expressed in murine host, where secondary anti-mouse antibodies may cause cross-reactivity. Binding is not Ca2+-dependent.
Form: Purified immunoglobulin in phosphate buffered saline plus 1% BSA and preservative. The conjugate protein concentration is approximately 1 mg/ml. |
• Immunocytochemistry
Working Dilution:
• 10 µg/ml by direct immunofluorescence using mammalian cells fixed with methanol:acetone |
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| Secondary Antibodies |
|
| Product # |
Package Size |
Product Name |
Characteristics |
Applications |
| A9044 |
2 ml |
Rabbit Anti-Mouse IgG, (whole molecule) Peroxidase Conjugate |
Specificity: Mouse IgG. Binds all mouse Igs.
Form: Solution in 0.01 M phosphate buffered saline, pH 7.4, containing 0.01% thimerosal as a preservative. |
Working Dilution:
• 1:6,000-8,000 by dot blotting
• 1:40,000 by direct ELISA
• 1:200 by immunohistochemistry (formalin-fixed,paraffin-embedded sections)
• 1:80,000 by indirect Western blotting (chemiluminescent) |
| A9917 |
1 ml |
Goat Anti-Mouse IgG, Fab Fragment Peroxidase Conjugate, Adsorbed with Human IgG |
Specificity: Mouse IgG Fab. Immunospecific purification removes essentially all goat serum proteins, including immunoglobulins, which do not specifically bind to the Fab fragment of mouse IgG. The antibody preparation is solid phase adsorbed with human IgG to ensure minimal cross reactivity in tissue or cell preparations.
Form: Solution in 0.01 M phosphate buffered saline, pH 7.4, containing 0.01% thimerosal as a preservative. |
Working Dilution:
• 1:8,000 by dot blotting
• 1:60,000 by direct ELISA
• 1:150 by immunohistochemistry (formalin-fixed,paraffin-embedded sections)
• 1:80,000 by indirect Western blotting (chemiluminescent) |
| A3682 |
1 ml |
Goat Anti-Mouse IgG, Fab Fragment Peroxidase Conjugate, Adsorbed with Human IgG and Rat Serum Proteins |
Specificity: Mouse IgG Fab. Immunospecific purification removes essentially all goatserum proteins, including immunoglobulins, which do not specifically bind to the Fab fragment of mouse IgG. The antibody preparation is solid phase adsorbed with human IgG and rat serum proteins to ensure minimal cross reactivity in tissue or cell preparations. Binds all Mouse Igs. |
Working Dilution:
• 1:4,000 by dot blotting
• 1:40,000 by direct ELISA
• 1:150 by immunohistochemistry (formalin-fixed,paraffin-embedded sections)
• 1:80,000 by indirect Western blotting (chemiluminescent) |
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| FLAG Western Blotting Kits |
|
| Product # |
Package Size |
Product Name |
Characteristics |
Applications |
| PQ0300 |
1 kit |
ProteoQwest™ FLAG® Colorimetric Western Blotting Kit, TMB Substrate |
Specificity: N-terminal, Met-N-terminal, Carboxy-terminal, or internal. Binding is not Ca2+-dependent.
Sensitivity: Colorimetric detection of as little as 1 ng of FLAG epitope-tagged fusion protein on Western blots using Monoclonal ANTI-FLAG® M2-Peroxidase (HRP) (Product Code A8592) and TMB substrate.
|
• Western blotting
Working Dilution:
• 1:1,000 by direct Western blotting (colorimetric) |
| PQ0400 |
1 kit |
ProteoQwest™ FLAG® Chemiluminescent Western Blotting Kit, CPS Substrate |
Specificity: N-terminal, Met-N-terminal, Carboxy-terminal, or internal. Binding is not Ca2+-dependent.
Sensitivity: Chemiluminescent detection of as little as 1 ng of FLAG epitope-tagged fusion protein on Western blots using Monoclonal ANTI-FLAG® M2-Peroxidase (HRP) (Product Code A8592) and CPS-1 substrate. |
• Western blotting
Working Dilution:
• 1:10,000 by direct Western blotting (chemiluminescent) |
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| FLAG Control Proteins |
|
| Product # |
Package Size |
Product Name |
Characteristics |
Applications |
| P5975 |
0.1 mg |
Met-FLAG® BAP Control Protein |
N-terminal Met-FLAG-BAP control protein is a 468 amino acid N-terminal Met-FLAG fusion protein of E. coli bacterial alkaline phosphatase (BAP)
MW: 49.4 kDa. |
Control protein for ANTI-FLAG M2 and M5 monoclonal antibodies in Western blotting, ELISA, immunoprecipitation, fluorescence microscopy, light microscopy, FACS and in immunoaffinity chromatography with the ANTI-FLAG M2 affinity gel. |
| P7457 |
0.1 mg |
Carboxy-terminal FLAG-BAP™ Control Protein |
A 466 amino acid C-terminal FLAG fusion protein of E. coli bacterial alkaline phosphatase (BAP).
MW: 49.1 kDa. |
Control protein for ANTI-FLAG M1 and M2 monoclonal antibodies in Western blotting, ELISA, immunoprecipitation, fluorescence microscopy, light microscopy, FACS, and in immunoaffinity chromatography |
| P7582 |
0.1 mg |
Amino-terminal FLAG-BAP™ Control Protein |
A 467 amino acid N-terminal FLAG fusion protein of E. coli bacterial alkaline phosphatase (BAP).
MW: 49.3 kDa. |
Control protein for the ANTI-FLAG M2 monoclonal antibody in immunological procedures such as Western blotting, ELISA, immunoprecipitation, fluorescence microscopy, light microscopy, FACS, and immunoaffinity chromatography. |
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| FLAG Plate |
|
| Product # |
Package Size |
Product Name |
Characteristics |
Applications |
| P2983 |
1 ea
|
ANTI-FLAG® High Sensitivity, M2 coated 96-well plates |
Specificity: N-terminal, Met-N-terminal or C-terminal of FLAG fusion proteins. Binding is not Ca2+-dependent.
Binding Capacity: Up to 300 ng/well.
Sensitivity: Detects 1 ng FLAG fusion protein/well by ELISA using p-NitrophenylPhosphate (pNPP) as a substrate.
Elution: 2X sample buffer, other elution buffers, or detect protein in the well without elution.
Form: The ANTI-FLAG®M2 antibody covalently attached through its Fc portion to the surface of a clear, polystyrene flat bottom plate. |
• Automation/High Throughput Screening for FLAG fusion protein expression
• ELISA
• Purification of FLAG fusion proteins
• Protein interaction screening (protein-protein or protein-organic) |
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