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.
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® M2 Antibody and is compatible with N-, C-terminal or internal tagging.
Immunoprecipitation (IP) / Co-IP
Mass spectrometry (MS)
ChIP/ RIP analysis
On-bead enzyme assays
Try out the DYKDDDDK IP Starter Pack (ffap)
DYKDDDDK Fab-Trap™ Agarose (20 reactions, product code ffa-20) for immunoprecipitation, 3xDYKDDDDK-peptide (1mg, fp-1) for elution and DYKDDDDK tag Polyclonal antibody (Binds to FLAG® tag epitope) (20 µL, 20543-1-AP-20UL) for detection in Western blots.
|Product DYKDDDDK Fab-Trap Agarose||Size 250 µL (10 reactions)||Code ffa-10||Price $ 275||Buy +|
|Product DYKDDDDK Fab-Trap Agarose||Size 500 µL (20 reactions)||Code ffa-20||Price $ 475||Buy +|
|Product DYKDDDDK Fab-Trap Agarose||Size 2.5 mL (100 reactions)||Code ffa-100||Price $ 2050||Buy +|
|Product DYKDDDDK Fab-Trap Agarose||Size 5 mL (200 reactions)||Code ffa-200||Price Please inquire|
|Product DYKDDDDK Fab-Trap Agarose||Size 10 mL (400 reactions)||Code ffa-400||Price Please inquire|
|Product DYKDDDDK Fab-Trap Agarose Kit||Size 500 µL (20 reactions)||Code ffak-20||Price $ 595||Buy +|
|Product 3xDYKDDDDK-peptide||Size 1 mg||Code fp-1||Price $ 105||Buy +|
|Product DYKDDDDK Immunoprecipitation Starter Pack||Size 20 reactions||Code ffap||Price $ 540||Buy +|
|Product Binding Control Agarose Beads||Size 500 µL (20 reactions)||Code bab-20||Price $ 75||Buy +|
|Product Spin Columns||Size 10 units||Code sct-10||Price $ 35||Buy +|
|Product Spin Columns||Size 20 units||Code sct-20||Price $ 60||Buy +|
|Product Spin Columns||Size 50 units||Code sct-50||Price $ 130||Buy +|
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
1x and 3xDYKDDDDK-tag sequence (1x and 3xFlag®-tag). The DYKDDDDK Fab-Trap™ Agarose binds to same epitope as Sigma's Anti-FLAG® M2 antibody.
25 µL slurry bind about 12.5 – 15 µg of recombinant 3xDYKDDDDK-tagged protein (~30 kDa)
Recombinant Fab fragment of monoclonal mouse IgG1 M2.1
Bead size: ~ 90 µm (cross-linked 4 % agarose beads)
Storage buffer: 1x PBS, Preservative: 0.09 % sodium azide
- SDS sample buffer
- 0.2 M glycine pH 2.5
Please consider whether elution is really needed. We recommend on-bead assays like on-bead digestion for MS analysis.
Compatibility with mass spectrometry
The DYKDDDDK Fab-Trap Agarose is optimized for on-bead digestion.
How to cite this product
Shipped at ambient temperature. Upon receipt store at 4°C. Stable for one year.
Should I use an N-terminal or C-terminal fusion tag?
Both the N-terminal or C-terminal fusion tag work well with the traps.
How can I avoid unspecific protein interactions binding to the trap?
How many mammalian cells are required for an immunoprecipitation reaction?
For one immunoprecipitation reaction, we recommend using ~10^6 - 10^7 mammalian cells. The yield is also dependent on the expression level of your protein of interest and the interaction partners.
How much cell extract should I use for an immunoprecipitation reaction?
For other type of cells than mammalian cells, we recommend using 0.5 - 1.0 mg of cell extract.
Do I need to elute bound proteins from the beads for mass spectrometry analysis?
No, you can directly conduct an on-bead digestion after immunoprecipitation. This procedure allows faster and more efficient sample preparation and a potential higher yield.
Do I need to elute my protein of interest from the beads for enzymatic assays?
What is the amount of trap slurry I need for one immunoprecipitation reaction?
25 µL slurry are sufficient for one pull-down reaction as the affinity of the DYKDDDDK Fab-Trap is very high.
DYKDDDDK Immunoprecipitation Starter Pack
You look for a cost-effective pack for Flag® IP or you are planning your first IP of a Flag-tagged protein? Or you want to test the DYKDDDDK Fab-Trap™ Agarose?
Here is a Starter Pack (ffap) containing
- DYKDDDDK Fab-Trap™ Agarose (20 reactions, ffa-20),
- 3xDYKDDDDK-peptide (1mg, fp-1) and
- DYKDDDDK tag Polyclonal antibody (Binds to FLAG® tag epitope) (20 µL, 20543-1-AP-20).
- Pulldown of Flag®-tagged proteins
- Lowest background in class
- Less contaminating antibody chains
- Reliable: recombinantly expressed Fab-fragment derived from M2 clone
- Small Fab-fragment tolerates harsh wash buffer conditions
- Convenient and fast: Easy handling and ready-to-use beads
- Highly selective binding of both N-and C-terminal Flag-tag
DYKDDDDK Fab-Trap Agarose Kit
The DYKDDDDK Fab-Trap Agarose is also available in a kit, including:
- DYKDDDDK Fab-Trap Agarose
- Lysis*, dilution and wash buffers
*The lysis buffer provided is suitable for mammalian cells. For other cell types use another suitable lysis buffer.
- Easy and convenient handling
- Rapid washing and clean elution of bound proteins
- Simplify the pulldown of your protein
- Spin columns for DYKDDDDK Fab-Trap Agarose
Binding Control for Agarose Beads
- Control for unspecific binding of proteins, DNA, etc. to beads
- Pre-clearing of cell lysate
- Binding control for DYKDDDDK Fab-Trap Agarose
Why is the DYKDDDDK Fab-Trap superior?
- Minimal contamination by Fab-fragment of the DYKDDDDK Fab-Trap
The DYKDDDDK Fab-Trap consists of a Fab-fragment, which is covalently bound to beads. In contrast to antibody-based immunoprecipitation reagents, there is no contamination from DYKDDDDK Fab-Trap on your SDS-PAGE and Western blot above 25 kDa, resulting in cleaner SDS-PAGE and Western blots. Moreover, you will find less antibody derived peptide contaminations in subsequent mass spectrometric analysis.
- Lowest Background in class
ChromoTek’s DYKDDDDK Fab-Trap is an anti-Flag affinity reagent based on a Fab-fragment, which is only a third of the size of a conventional antibody. Due to its smaller size, the Fab-fragment minimizes unwanted interaction with the host-cell proteome, resulting in efficient immunoprecipitations with superior purity and less background.
- Small Fab-fragment tolerates harsh wash buffer conditions
The small Fab-fragment of the DYKDDDDK Fab-Trap tolerates harsher washing conditions as comparable anti-Flag antibody affinity resins. The possibility to use harsher wash buffers, enables effective removal of unwanted cellular proteins for highest purity immunoprecipitation.
- Tight and highly selective binding of a single Flag-tag
The DYKDDDDK Fab-Trap binds with high specificity to N- or C-terminal Flag-tagged proteins. In addition, it has a high affinity to both 1x or 3xFlag-tag enabling a broad experimental applicability.
- Ready-to-use reagent
DYKDDDDK Fab-Trap is a ready-to-use reagent that comes pre-conjugated to beads. There is no need for tedious and time-consuming pre-incubation steps required for pulldowns with protein A or G beads making the immunoprecipitation faster, more convenient, and easier to reproduce.
- Mild, competitive elution with DYKDDDDK-peptide or acidic conditions
DYKDDDDK Fab-Trap can be mildly eluted with 3xDYKDDDDK-peptide, preserving the native structure of your Flag-tagged protein for structural and activity downstream analysis. In addition, convenient elution by low pH or SDS-sample buffer offers fast and efficient elution performance.
- Reproducible results
DYKDDDDK Fab-Trap binds to the same epitope as Sigma’s ANTI-FLAG® M2 antibody. The Fab-fragment of the DYKDDDDK Fab-Trap is recombinantly produced for the generation of highly reliable data.
Only for research applications, not for diagnostic or therapeutic use!