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GFP-Trap Magnetic Particles M-270: Most cited GFP-Nanobody conjugated to Magnetic Particles M-270

Posted by Christoph Eckert on Jan 17, 2020 11:27:30 AM

GFP-Trap® Magnetic Particles M-270 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 Magnetic Particles M-270. Hence, also GFP-Trap Magnetic Particles M-270 has the very high affinity of 1 pM like GFP-Trap Agarose and Magnetic Agarose.


GFP-Trap Dynabeads_Coomassie_WB groß_2Immunoprecipitation of GFP with GFP-Trap Magnetic Particles M-270.
I: Input, FT: Flow-Through, B: Bound

ChromoTek® GFP-Trap Magnetic Particles M-270 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 Magnetic Particles M-270
  • automation and high throughput applications

GFP-Trap Magnetic Particles M-270 complements GFP-Trap Magnetic Agarose and GFP-Trap Agarose.

GFP-Trap Magnetic Particles M-270 for IP of large GFP-fusion proteins and protein complexes
Both magnetic agarose and agarose are porous beads, whereas Magnetic Particles M-270 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.
Magnetic Particles M-270 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 Magnetic Particles M-270 and are effectively immunoprecipitated.

The GFP-Trap Magnetic Particles M-270 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 Magnetic Particles M-270 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

Nano-Trap Magnetic Agarose vs. Magnetic Particles 2021Cartoon 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 Magnetic Particles M-270.


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 Magnetic Particles M-270 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 Magnetic Particles M-270 and the efficiency of the IP is high.

See for yourself and test the GFP-Trap Magnetic Particles M-270:

Request GFP-Trap Magnetic Particles M-270 sample

 

Here are some FAQs:

Can I re-use GFP-Trap Magnetic Particles M-270?
Re-use or multiple use of GFP-Trap Magnetic Particles M-270 is not recommended as SDS is required for complete elution.

How should I store GFP-Trap Magnetic Particles M-270?
GFP-Trap Magnetic Particles M-270 should be stored at + 4°C/ +40°F. Do not freeze GFP-Trap Magnetic Particles M-270.

What are GFP-Trap Magnetic Particles M-270 made of?
GFP-Trap Magnetic Particles M-270 consists of ChromoTek’s GFP Nanobody and Magnetic Particles M-270. Magnetic Particles M-270 are magnetic spherical polymer particles and have a solid surface.

How big are GFP-Trap Magnetic Particles M-270?
The medium particle size of GFP-Trap Magnetic Particles M-270 is 2.8 µm.

How much GFP-Trap Magnetic Particles M-270 should I use for an IP?
We recommend 25 μL of bead slurry of GFP-Trap Magnetic Particles M-270.

How to remove GFP-Trap Magnetic Particles M-270?
GFP-Trap Magnetic Particles M-270 are magnetic and can be removed by using a magnet or magnetic stand.

GFP-Trap Magnetic Particles M-270 consists of the most cited GFP Nanobody conjugated to Magnetic Particles M-270.

White papers discussing various aspects of GFP-Trap performance: 

Topics: GFP Immunoprecipitation, GFP Nanobody

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