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SAB4200071 Sigma-Aldrich

ANTI-FLAG® antibody, Rat monoclonal

clone 6F7, purified from hybridoma cell culture

Synonym: Anti-ddddk, Anti-dykddddk

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Properties

Related Categories Alphabetical Index, Antibodies, Antibodies and Conjugates, Antibodies to Peptide and Protein Fusion Tags, FK-FO,
conjugate   unconjugated
clone   6F7, monoclonal
biological source   rat
application(s)   immunoprecipitation (IP): 2.5-5.0 μg using lysates of transiently transfected cells expressing C-terminal-FLAG-tagged protein
  western blot: 0.5-1.0 μg/mL using extracts of transiently transfected cells expressing C-terminal-FLAG-tagged protein
species reactivity   all
form   buffered aqueous solution
shipped in   dry ice
storage temp.   −20°C
antibody form   purified from hybridoma cell culture
  purified immunoglobulin
isotype   IgG1
antibody product type   primary antibodies
immunogen sequence   (DYKDDDDK)

Description

General description

Monoclonal Anti-FLAG® (rat IgG1 isotype) is derived from the hybridoma 6F7 produced by the fusion of mouse myeloma cells and splenocytes from rat immunized with the FLAG® peptide. The antibody is purified from culture supernatant of hybridoma cells grown in a bioreactor.

Monoclonal Anti-FLAG® recognizes N-terminal,
C-terminal and internal Flag-tagged fusion proteins. The product is especially recommended for identifying C-terminal FLAG®-tagged fusion proteins. The antibody may be used in various immunochemical techniques including immunoblotting and immunoprecipitation.

Epitope tags provide a method to localize gene products in a variety of cell types, study the topology of proteins and protein complexes, identify associated proteins, and characterize newly identified, low abundance, or poorly immunogenic proteins when protein specific antibodies are not available. Tagging with the FLAG® peptide sequence may be done at the N-terminus, N-terminus preceded by a methionine residue, C-terminus, or at internal positions of the target protein. FLAG may also be placed in associationith other tags. The small size of the FLAG® tag or sequence and its high hydrophilicity tend to decrease the possibility of interference with the protein expression, proteolytic maturation, antigenicity, and function.

The N-terminal FLAG® peptide sequence contains a unique enterokinase cleavage site allowing it to be completely removed from the purified fusion proteins. Cleavage of the C-terminal FLAG® peptide from a fusion protein catalyzed by Cu2+ ions has been reported. A sequence motif with five out of eight amino acid residues identical to the FLAG peptide is found in both rat and mouse Mg2+dependent protein b-phosphatase, as well as in the human and bovine enzyme.

Immunogen

FLAG peptide

DYKDDDDK

Application

Browse additional application references in our FLAG® Literature portal.

Monoclonal Anti-FLAG (rat IgG1 isotype) recognizes N-terminal, C-terminal and internal Flag-tagged fusion proteins. The product is especially recommended for identifying C-terminal FLAG-tagged fusion proteins.

Physical form

Solution in 0.01 M phosphate buffered saline, pH 7.4, containing 15 mM sodium azide.

Legal Information

FLAG is a registered trademark of Sigma-Aldrich Co. LLC

Safety & Documentation

Safety Information

RIDADR 
NONH for all modes of transport

Documents

Certificate of Analysis

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Protocols & Articles

Articles

Antibody Basics

Immunoglobulins (Igs) are produced by B lymphocytes and secreted into plasma. The Ig molecule in monomeric form is a glycoprotein with a molecular weight of approximately 150 kDa that is shaped more ...
Keywords: Affinity chromatography, Centrifugation, Chromatography, Digestions, Direct immunofluorescence, Gene expression, High performance liquid chromatography, Immunofluorescence, Ion Exchange, Microscopy, Precipitation, Purification, Rheumatology, Scanning electron microscopy

Protocols

Western Blot Protocol | Immunoblotting Protocol

Western Blotting refers to the electrophoretic transfer of proteins from sodium dodecyl sulfate polyacrylamide gels to sheets of PVDF or nitrocellullose membrane, followed by immunodetection of prote...
Keywords: AGE, Buffers, Cell disruption, Detergents, Dialysis, Electroblotting, Electrophoresis, Enzyme activity, Gel electrophoresis, Immunoprecipitation, PAGE, Protein extraction, Purification, Sample preparations, Western blot

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Peer-Reviewed Papers
15

References

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