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SAB4200071

Sigma-Aldrich

ANTI-FLAG® antibody, Rat monoclonal

clone 6F7, purified from hybridoma cell culture

Synonym(s):

Anti-ddddk, Anti-dykddddk

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About This Item

MDL number:
MFCD02262912
UNSPSC Code:
12352203
NACRES:
NA.32

biological source

rat

conjugate

unconjugated

antibody form

purified from hybridoma cell culture
purified immunoglobulin

antibody product type

primary antibodies

clone

6F7, monoclonal

form

buffered aqueous solution

species reactivity

all

technique(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

isotype

IgG1

immunogen sequence

(DYKDDDDK)

shipped in

dry ice

storage temp.

−20°C

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.
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

ANTI-FLAG® antibody, Rat monoclonal has been used in:
  • chromatin immunoprecipitation (ChIP)
  • western blotting
  • coimmunoprecipitation
  • flow cytometric analysis

Learn more product details in our FLAG® application portal.
Browse additional application references in our FLAG® Literature portal.

Physical form

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

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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Storage Class

10 - Combustible liquids

flash_point_f

Not applicable

flash_point_c

Not applicable

Certificates of Analysis (COA)

Search for Certificates of Analysis (COA) by entering the products Lot/Batch Number. Lot and Batch Numbers can be found on a product’s label following the words ‘Lot’ or ‘Batch’.

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PHF21B overexpression promotes cancer stem cell-like traits in prostate cancer cells by activating the Wntbeta-catenin signaling pathway
Li Q, et al
Journal of Experimental & Clinical Cancer Research, 36(1), 85-85 (2017)
Primary resistance mechanism of the canine distemper virus fusion protein against a small-molecule membrane fusion inhibitor
Kalbermatter D, et al.
Virus Research, 259, 28-37 (2019)
Rotaviral enterotoxin nonstructural protein 4 targets mitochondria for activation of apoptosis during infection
Bhowmick R, et al.
The Journal of Biological Chemistry, 287(42), 35004-35020 (2012)
D P Humphreys et al.
Protein engineering, 12(2), 179-184 (1999-04-09)
The peptide sequence (N)DKTH(C) was investigated as a site for efficient, specific cleavage of a fusion protein by cupric ions using a humanised gamma1 Fab' as a model protein. The native upper hinge (N)DKTH(C) sequence was mutated to create a
A S Robeva et al.
Biochemical pharmacology, 51(4), 545-555 (1996-02-23)
An expression plasmid for mammalian cells (CLDN10B) has been modified to add nucleotides encoding hexahistidine and the FLAG peptide (H/F) to cDNAs. The new mammalian expression plasmid has been named pDoubleTrouble (pDT). The plasmid and a recombinant baculovirus were used
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