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ChromoTek DYKDDDDK Fab-Trap™ Agarose

The ChromoTek DYKDDDDK Fab-Trap™ Agarose consists of an anti-DYKDDDDK-tag Fab-fragment, which is covalently bound to agarose beads. DYKDDDDK Fab-Trap™ Agarose is used to immunoprecipitate 1x and 3xDYKDDDDK-tagged fusion proteins from cell extracts of various organisms like mammals, plants, bacteria, yeast, insects etc.

Specificity

Flag®-tag (1x and 3x)

Applications

IP, CoIP, ChIP, RIP, Protein purification

Type

Fab-fragment

Conjugate

Agarose

Cat no : ffa

Synonyms

Fab-Trap Agarose, Fab-Trap, DYKDDDDK, flag

Validation Data Gallery

  • DYKDDDDK Fab-Trap™ was used for immunoprecipitation of Flag®-tagged protein from HEK293T cell lysate. During elution with 3xDYKDDDDK-peptide (fp-1) the enriched Flag® fusion protein is released from DYKDDDDK Fab-Trap™. Western blot was probed with DYKDDDDK tag Polyclonal antibody (Binds to FLAG® tag epitope) (Proteintech, 20543-1-AP) and Nano-Secondary® alpaca anti-human IgG/anti-rabbit IgG, recombinant VHH, Alexa Fluor® 488 [CTK0101, CTK0102] (srbAF488-1). L: Prestained protein marker (Proteintech, PL00001), I: Input, FT: Flow-Through, E: Elution.

    DYKDDDDK Fab-Trap™ was used for immunoprecipitation of Flag®-tagged protein from HEK293T cell lysate. During elution with 3xDYKDDDDK-peptide (fp-1) the enriched Flag® fusion protein is released from DYKDDDDK Fab-Trap™. Western blot was probed with DYKDDDDK tag Polyclonal antibody (Binds to FLAG® tag epitope) (Proteintech, 20543-1-AP) and Nano-Secondary® alpaca anti-human IgG/anti-rabbit IgG, recombinant VHH, Alexa Fluor® 488 [CTK0101, CTK0102] (srbAF488-1). L: Prestained protein marker (Proteintech, PL00001), I: Input, FT: Flow-Through, E: Elution.

  • DYKDDDDK Fab-Trap™ was used for immunoprecipitation of Flag®-tagged protein from HEK293T cell lysate. During elution with 2x SDS-sample buffer the enriched Flag® fusion protein and Fab-fragment are released from DYKDDDDK Fab-Trap™. Western blot was probed with DYKDDDDK tag Polyclonal antibody (Binds to FLAG® tag epitope) (Proteintech, 20543-1-AP) and Nano-Secondary® alpaca anti-human IgG/anti-rabbit IgG, recombinant VHH, Alexa Fluor® 488 [CTK0101, CTK0102] (srbAF488-1). L: Prestained protein marker (Proteintech, PL00001), I: Input, FT: Flow-Through, B: Bound.

    DYKDDDDK Fab-Trap™ was used for immunoprecipitation of Flag®-tagged protein from HEK293T cell lysate. During elution with 2x SDS-sample buffer the enriched Flag® fusion protein and Fab-fragment are released from DYKDDDDK Fab-Trap™. Western blot was probed with DYKDDDDK tag Polyclonal antibody (Binds to FLAG® tag epitope) (Proteintech, 20543-1-AP) and Nano-Secondary® alpaca anti-human IgG/anti-rabbit IgG, recombinant VHH, Alexa Fluor® 488 [CTK0101, CTK0102] (srbAF488-1). L: Prestained protein marker (Proteintech, PL00001), I: Input, FT: Flow-Through, B: Bound.

  • Analysis of unspecific binding of endogenous proteins to the DYKDDDDK Fab-Trap™ in the absence of Flag® fusion protein. Lysates devoid of Flag® fusion protein from HEK cells (HEK293T), Yeast (Saccharomyces cerevisiae), Insect cells (HighFive), Bacteria (Escherichia coli TG1) , and Plant (Arabidopsis thaliana) were subjected to immunoprecipitation with DYKDDDDK Fab-Trap™ and elution with 2x SDS-sample buffer. SDS-PAGE analysis shows elution of Fab-fragment only without contamination of endogenous proteins. L: Prestained protein marker (Proteintech, PL00001), I: Input, B: Bound.

    Analysis of unspecific binding of endogenous proteins to the DYKDDDDK Fab-Trap™ in the absence of Flag® fusion protein. Lysates devoid of Flag® fusion protein from HEK cells (HEK293T), Yeast (Saccharomyces cerevisiae), Insect cells (HighFive), Bacteria (Escherichia coli TG1) , and Plant (Arabidopsis thaliana) were subjected to immunoprecipitation with DYKDDDDK Fab-Trap™ and elution with 2x SDS-sample buffer. SDS-PAGE analysis shows elution of Fab-fragment only without contamination of endogenous proteins. L: Prestained protein marker (Proteintech, PL00001), I: Input, B: Bound.

Product Information

The ChromoTek DYKDDDDK Fab-Trap™ Agarose consists of an anti-DYKDDDDK-tag Fab-fragment, which is covalently bound to agarose beads. DYKDDDDK Fab-Trap™ Agarose is used to immunoprecipitate 1x and 3xDYKDDDDK-tagged fusion proteins from cell extracts of various organisms like mammals, plants, bacteria, yeast, insects etc.

DescriptionThe DYKDDDDK Fab-Trap Agarose is a premium affinity resin for efficient immunoprecipitation (IP) of Flag®-tagged proteins. The DYKDDDDK Fab-Trap Agarose consists of a Fab-fragment, which is covalently bound to agarose beads.

• Lowest background in class

• Less contaminating antibody chains

• Recombinantly expressed Fab-fragment derived from M2 clone

• Compatible with harsh buffer conditions

• Ready-to-use

ApplicationsIP, CoIP, ChIP, RIP, Protein purification
Specificity/TargetThe DYKDDDDK Fab-Trap™ Agarose specifically binds to 1x and 3xDYKDDDDK-tag sequence (1x and 3xFlag®-tag). It binds to same epitope as Sigma's Anti-FLAG® M2 Antibody and is compatible with N-, C-terminal or internal tagging.
Binding capacity12.5 - 15 µg of recombinant 3xDYKDDDDK-tagged protein (~30 kDa) per 25µL bead slurry
ConjugateAgarose beads; bead size: ~ 90 µm (cross-linked 4 % agarose beads)
Elution buffer3xDYKDDDDK-peptide
SDS sample buffer
0.2 M glycine pH 2.5
Wash buffer compatibility0.5 % Nonidet P40 Substitute, 5 % Tween™ 20, 2 M NaCl, 10 mM TCEP, 10 mM DTT, 10 mM ß-mercaptoethanol, 2 M Urea
Type Fab-fragment
ClassRecombinant
HostMouse
RRIDAB_2894836
Storage Buffer1x PBS, Preservative: 0.09 % sodium azide
Storage ConditionShipped at ambient temperature. Upon receipt store at 4°C. Stable for one year. Do not freeze!

Publications

ApplicationTitle
IP

J Mol Biol

The integration of proteome-wide PTM data with protein structural and sequence features identifies phosphorylations that mediate 14-3-3 interactions

Authors - Ce Egbert
View Article
IP

Cell Rep

SPATA2 restricts OTULIN-dependent LUBAC activity independently of CYLD

Authors - Laura Griewahn
View Article

Reviews

The reviews below have been submitted by verified Proteintech customers who received an incentive forproviding their feedback.

FH

Jonasz (Verified Customer) (11-15-2022)

The ffa-2 DYKDDDDK Fab-Trap agarose fulfilled our expection and showed excellent performance comparable to other Trap products by Chromotek/Proteintech such as GFP-Trap or myc-Trap agarose. We successfully precipitated FLAG-tagged ubiquitin overexpressed in HEK293T cells.

  • Applications: Immunoprecipitation
  • Cell Tissue Type: HEK 293T cells
FH

Abida (Verified Customer) (11-02-2022)

Stain-free gel (A) and western blot (B) of immunoprecipitation of flag-fusion protein from Lactococcus lactis bacterial cell lysate with DYKDDDDK Fab trap agarose beads (Proteintech, cat no ffa). Flag-fusion protein was eluted from DYKDDDDK Fab-Trap with 2x SDS-sample buffer. Western blot was probed with rabbit DYKDDDDK tag polyclonal antibody (Proteintech, cat no 20543-1-AP) followed by anti-rabbit StarBright IgG Blue 520 fluorescent goat secondary antibody (BioRad, cat no 12005869). I: input (lysate), M: protein marker, FT: flowthrought, B: bound (elution).

  • Applications: Immunoprecipitation
  • Primary Antibody Dilution: 1:5000
  • Cell Tissue Type: Bacteria Lactococcus lactis
FH

Tobias (Verified Customer) (10-25-2022)

excellent efficiency and yield.

  • Applications: Immunoprecipitation
FH

Peter (Verified Customer) (10-24-2022)

Excellent recovery of flag tagged proteins with short incubation times

  • Applications: Immunoprecipitation
  • Cell Tissue Type: A549
FH

Amit (Verified Customer) (03-16-2022)

Works well for purification of FLAG tag protein purification and immunoprecipitation. Highly recommended!

  • Applications: Immunoprecipitation,Protein purification
  • Cell Tissue Type: E.Coli