ChromoTek DYKDDDDK Fab-Trap® Agarose

Validation Data Gallery

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  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.

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  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.

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  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.

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.

Cat No. ffa

Publications(44)

Reviews (12)

Specificity

Flag®-tag (1x and 3x)

Applications

IP, CoIP, ChIP, RIP, Protein purification

Type

Fab-fragment

Conjugate

Agarose

Synonyms

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

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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.

• Description: The 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;

  • Compatible with harsh buffer conditions;

  • Ready-to-use

• Applications: IP, CoIP, ChIP, RIP, Protein purification
• Specificity/Target: The 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® Antibody and is compatible with N-, C-terminal or internal tagging.
• Binding capacity: 12.5 - 15 µg of recombinant 3xDYKDDDDK-tagged protein (~30 kDa) per 25µL bead slurry
• Conjugate: Agarose beads; bead size: ~ 90 µm (cross-linked 4 % agarose beads)
• Elution buffer: 3xDYKDDDDK-peptide; SDS sample buffer; 0.2 M glycine pH 2.5
• Wash buffer compatibility: 0.5 % 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
• Class: Recombinant
• Host: Mouse
• RRID: AB_2894836
• Storage Buffer: 1x PBS, Preservative: 0.09 % sodium azide
• Storage Condition: Shipped at ambient temperature. Upon receipt store at 4°C. Stable for one year. Do not freeze!
• Size: 25ul/reactions (eg：20rxns=500ul slurry)

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Publications

Application	Title
IP	EMBO Mol Med; Targeting PI3K/Akt/mTOR signaling in rodent models of PMP22 gene-dosage diseases; Authors - Doris Krauter
CoIP	Cell Rep; LHP1 and INO80 cooperate with ethylene signaling for warm ambient temperature response by activating specific bivalent genes; Authors - Zhengyao Shao
IP	mBio; The Cajal body protein p80-coilin forms a complex with the adenovirus L4-22K protein and facilitates the nuclear export of adenovirus mRNA; Authors - Laura White
CoIP	Open Biol; Coiled-coil-mediated dimerization of Atg16 is required for binding to the PROPPIN Atg21; Authors - Miranda Bueno-Arribas
IP	J Biol Chem; ZNF692 regulates nucleolar morphology by interacting with NPM1 and modifying its self-assembly properties; Authors - Isabella N Brown
IP	Development; Contributions of the Dachsous intracellular domain to Dachsous-Fat signaling; Authors - Bipin Kumar Tripathi

Reviews

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

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FH

Arpita (Verified Customer) (04-15-2025)

NA

• Applications: Protein purification

FH

Nicolas (Verified Customer) (04-03-2025)

I used the DYKDDDDK fab trap agarose beads to perform immunoprecipitation of my protein of interest. It worked very well and did not require high amount of material. The possibility to elute in 3 different way is also a plus regarding what you want to do with your protein afterward.

• Applications: Immunoprecipitation
• Cell Tissue Type: Bacteria, Caulobacter crescentus

FH

Elizabeth (Verified Customer) (03-10-2025)

The DYKDDDDK Fab-Trap Agarose has high affinity for my FLAG-tagged protein!

• Applications: Immunoprecipitation,Protein purification
• Cell Tissue Type: Chinese hamster ovary cells

FH

Jianhua (Verified Customer) (02-20-2025)

Great, works well for IP

• Applications: Immunoprecipitation
• Cell Tissue Type: 293

FH

Hanna (Verified Customer) (11-19-2024)

The immunoprecipitation of FLAG-tagged protein to study its interactome using the Fab-trap was so effective and easy to perform

• Applications: Immunoprecipitation
• Cell Tissue Type: Neuroblastoma cell line

FH

Alexander (Verified Customer) (08-29-2024)

Here we have tested the ChromoTek DYKDDDDK Fab-Trap Agarose beads (Cat No. ffa) to detect a protein of interest, which is expressed as a fusion protein with a tandem-peptide tag consisting of a Twin-Streptag and a FLAG-tag. In the attached image, we have pulled down the FLAG-tagged fusion protein from cortical organoid lysates. For the western blot we used TBS-T + 1% BSA for blocking (1h at RT) and TBS-T + 5 % BSA for diluting the antibodies. The FLAG-fusion protein was detected with a mouse-anti-FLAG M2 monoclonal antibody. The immunoprecipitation of the FLAG-tagged fusion protein with the DYKDDDDK Fab-Trap Agarose beads showed excellent efficiency as well as protein yield and does not show unspecific binding in the wildtype control samples.

• Applications: Western Blot,Immunoprecipitation
• Primary Antibody Dilution: 1:1000
• Cell Tissue Type: cortical organoids

FH

Hana (Verified Customer) (06-08-2023)

In this immunoprecipitation experiment, I have tested your DYKDDDDK Fab-Trap™ Agarose (ffa) beads, which are very nicely and effectively pulling down FLAG-tagged MCMBP protein. MCMBP is a strong interactor of MCMs. In the attached image, we have pulled down the MCM7 and MCM5 subunits as we previously described in Sedlackova et. al., Nature, 2020. Additionally in this experiment, we tested the FLAG antibody (80010-1-RR, dilution for western blot was 1:1000, 5%milk in 1xPBS+0,1% Tween, incubated overnight) and MCM5 antibody (11703-1-AP, dilution for western blot was 1:1000, 5%milk in 1xPBS+0,1% Tween, incubated overnight). All your products worked amazingly well.

• Applications: Western Blot,Immunoprecipitation
• Cell Tissue Type: human osteosarcoma U2OS

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