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F2555

Sigma-Aldrich

Monoclonal ANTI-FLAG® antibody produced in rabbit

clone SIG1-25, ascites fluid

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Synonym(s):
Anti-ddddk, Anti-dykddddk
NACRES:
NA.32

biological source

rabbit

Quality Level

conjugate

unconjugated

antibody form

ascites fluid

antibody product type

primary antibodies

clone

SIG1-25, monoclonal

technique(s)

immunocytochemistry: 1:125-1:250 using transiently transfected cells expressing FLAG (sequence at the N-terminus)-tagged protein fixed with paraformaldehyde/Triton X-100
indirect ELISA: suitable
western blot: 1:250-1:500 using extracts of transiently transfected cells expressing FLAG (sequence at the N-terminus)-tagged protein

isotype

IgG

immunogen sequence

DYKDDDDK

shipped in

dry ice

storage temp.

−20°C

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F7425F3040F3165
conjugate

unconjugated

conjugate

unconjugated

conjugate

unconjugated

conjugate

unconjugated

antibody form

ascites fluid

antibody form

affinity isolated antibody

antibody form

purified immunoglobulin

antibody form

purified immunoglobulin (Purified IgG1 subclass)

clone

SIG1-25, monoclonal

clone

polyclonal

clone

M1, monoclonal

clone

M2, monoclonal

technique(s)

immunocytochemistry: 1:125-1:250 using transiently transfected cells expressing FLAG (sequence at the N-terminus)-tagged protein fixed with paraformaldehyde/Triton X-100, indirect ELISA: suitable, western blot: 1:250-1:500 using extracts of transiently transfected cells expressing FLAG (sequence at the N-terminus)-tagged protein

technique(s)

dot blot: 1-2.5 μg/mL, immunoprecipitation (IP): 4-8 μg using amino terminal FLAG-BAP fusion protein from E. coli crude lysate, indirect immunofluorescence: 5-10 μg/mL using 293T cells transfected with a plasmid encoding FLAG-JNK, western blot (chemiluminescent): 1-2.5 μg/mL using an E. coli periplasmic extract expressing an N-terminal FLAG fusion protein

technique(s)

western blot: 10 μg/mL

technique(s)

western blot: 10 μg/mL (Protein A)

isotype

IgG

isotype

IgG

isotype

-

isotype

IgG1

General description

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 association with 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 catalyzed by Cu2+ ions of the C-terminal FLAG® peptide from a fusion protein 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. This rabbit ANTI-FLAG® antibody is produced from the hybridoma SIG1-25. The hybridoma is derived by the fusion of myeloma cells and splenocytes from rabbits immunized with the FLAG® peptide sequence conjugated to KLH.

Specificity

This monoclonal antibody recognizes the FLAG sequence at the N-terminus

Immunogen

FLAG peptide sequence DYKDDDDK

Application

Monoclonal ANTI-FLAG® antibody produced in rabbit has been used in:
  • immunofluorescence
  • western blot analysis
  • immunocytochemistry
  • indirect ELISA

Browse additional application references in our FLAG® Literature portal.

Physical form

Provided as ascites fluid containing 15 mM sodium azide as a preservative.

Legal Information

ANTI-FLAG is a registered trademark of Merck KGaA, Darmstadt, Germany
FLAG is a registered trademark of Merck KGaA, Darmstadt, Germany
Triton is a trademark of The Dow Chemical Company or an affiliated company of Dow

Storage Class Code

10 - Combustible liquids

WGK

WGK 3

Flash Point(F)

Not applicable

Flash Point(C)

Not applicable

Personal Protective Equipment

dust mask type N95 (US), Eyeshields, Gloves

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