GFP-Trap® Dynabeads is optimized for the immunoprecipitation (IP) of large GFP-tagged proteins and for Co-IP of protein complexes. It consists of ChromoTek’s established anti-GFP Nanobody conjugated to Dynabeads™. Hence, also GFP-Trap Dynabeads has the very high affinity of 1 pM like GFP-Trap Agarose and Magnetic Agarose.
Immunoprecipitation of GFP with GFP-Trap Dynabeads.
I: Input, FT: Flow-Through, B: Bound
ChromoTek® GFP-Trap Dynabeads can be used for
- IP and Co-IP of GFP-tagged proteins of all kind of sizes and shapes, e.g. Megadalton complexes
- magnetic separation with easy and efficient washing of Dynabeads
- automation and high throughput applications
GFP-Trap Dynabeads for IP of large GFP-fusion proteins and protein complexes
Both magnetic agarose and agarose are porous beads, whereas Dynabeads are solid. The pore size of GFP-Trap Agarose and Magnetic Agarose can be too small for large GFP-fusion proteins, multimers, or bulky complexes with binding partners to diffuse into the pores and to interact with the majority of the GFP VHH ligands. Next to size also the protein’s shape can limit diffusion. Therefore, binding can just occur on the small fraction of GFP-VHH ligands that are conjugated to the outer surface of the GFP-Trap Agarose or Magnetic Agarose beads. This results in a poor immunoprecipitation performance.
Dynabeads are non-porous; ligands are coupled on their surface. Hence, all size GFP-tagged proteins including very large GFP-tagged proteins, oligomers and complexes including bulky interaction partners bind to all GFP VHH ligands of GFP-Trap Dynabeads and are effectively immunoprecipitated.
The GFP-Trap Dynabeads is optimized for the immunoprecipitation of
- very large GFP-fusion proteins
- GFP-fusion protein multimers like dimers, tetramers, etc.
- GFP-fusion proteins that are part of bulky protein complexes
Furthermore, GFP-Trap Dynabeads is optimized for the Co-IP of
- GFP-fusion proteins binding to bulky interaction partners
- GFP-fusion proteins with multiple binding partners that form large complexes
Cartoon to visualize the binding of large GFP-fusion proteins (GFP (green) + protein of interest (POI, blue) with interacting partner X (Prot X, orange) or multimeric proteins (GFP (green) + protein of interest (POI, blue) to the GFP VHH (dark green) of GFP-Trap Magnetic Agarose or GFP-Trap Dynabeads.
Co-immunprecipitation of a protein complex:
The large complex of GFP-fusion protein and interaction partner is too large to diffuse into the pores of magnetic agarose beads and therefore binds only to the small fraction of GFP VHHs on the outer surface. Most of the complex of GFP-fusion protein and interaction partner is washed away, which results in a poor performance. In contrast, the GFP-Trap Dynabeads bind all GFP-fusion protein complexes. Bound GFP-fusion protein complexes are not removed during washing, which leads to high performance IPs.
Immunoprecipitation of a multimeric protein:
Some proteins tend to form multimeric proteins such as dimers or tetramers. The shown tetramer is too large to diffuse into the pores of magnetic agarose beads and only few multimeric proteins are bound. On the other hand, all multimeric proteins are bound by GFP-Trap Dynabeads and the efficiency of the IP is high.
Here are some FAQs:
Can I re-use GFP-Trap Dynabeads?
Re-use or multiple use of GFP-Trap Dynabeads is not recommended as SDS is required for complete elution.
How should I store GFP-Trap Dynabeads?
GFP-Trap Dynabeads should be stored at + 4°C/ +40°F. Do not freeze GFP-Trap Dynabeads.
What are GFP-Trap Dynabeads made of?
GFP-Trap Dynabeads consists of ChromoTek’s GFP Nanobody and Dynabeads M-270. Dynabeads M-270 are magnetic spherical polymer particles and have a solid surface.
How big are GFP-Trap Dynabeads?
The medium particle size of GFP-Trap Dynabeads is 2.8 µm.
How much GFP-Trap Dynabeads should I use for an IP?
We recommend 25 μL of bead slurry of GFP-Trap Dynabeads.
How to remove GFP-Trap Dynabeads?
GFP-Trap Dynabeads are magnetic and can be removed by using a magnet or magnetic stand.
GFP-Trap Dynabeads consists of the most cited GFP Nanobody conjugated to Dynabeads, the most cited beads in science.
White papers discussing various aspects of GFP-Trap performance: