ChromoTek GFP-Trap® is optimized for the immunoprecipitation of GFP-tagged proteins and their interacting factors.
The GFP-Trap consists of the ChromoTek anti-GFP Nanobody/ VHH that is coupled to 3 different types of beads or is immobilized in a 96 multiwell plate:
- Agarose beads
- Magnetic Agarose beads
- Magnetic Particles M-270
- 96 Multiwell Plate
Immunoprecipitation with GFP-Trap.
I: Input, FT: Flow-Through, B: Bound
Based on the properties of the different matrices (see table), we recommend:
- GFP-Trap Agarose for very low background and high binding capacity IP
- GFP-Trap Magnetic Agarose for magnetic separation and high binding capacity IP
- GFP-Trap Magnetic Particles M-270 for IP of very large proteins/complexes
- GFP-Trap Multiwell Plates for high throughput analysis
|
GFP-Trap |
Agarose |
Magnetic Agarose |
Magnetic Particles M-270 |
Multiwell Plate |
|
Low background* |
+++ |
++ |
++ |
++ |
|
Binding capacity |
+++ |
+++ |
+ |
+ |
|
Size GFP-tagged protein* |
Small to large |
Small to large |
Small to very large |
Small to very large |
|
Bead separation |
centrifugation |
magnetic |
magnetic |
Nanobody directly immobolised to multiwell plate |
*Please note that these are general guidelines. For a specific protein or complex, the optimal experimental conditions/ GFP-Trap have to be tested.
In detail, all types of GFP-Trap have very low background binding due to the highly specific binding of GFP (fusion proteins). In comparison, GFP-Trap Agarose has the lowest background. The background of GFP-Trap Magnetic Agarose and GFP-Trap Magnetic Particles M-270 is at slightly elevated levels. Hence, GFP-Trap Agarose is recommended if a very low background is needed.
The size and shape of the actual GFP-fusion protein, multimer or complex may require the use of the non-porous GFP-Trap Magnetic Particles M-270, because larger GFP-fusion proteins can’t access the GFP VHH ligands inside the pores of agarose and magnetic agarose beads. Note that smaller proteins may form multimers or may be part of larger complexes and hence have larger sizes than expected.
Transparent GFP-Trap Multiwell Plates are pre-blocked and the GFP Nanobody is immobilized within the wells on solid polystyrene surface. They can be used for for high throughput analysis and immunoprecipitation of even very large proteins/ protein complexes.
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|
GFP-Trap beads |
||
|
|
Agarose |
Magnetic Agarose |
Magnetic Particles M-270 |
|
Matrix |
Agarose |
Magnetic agarose |
Magnetic Particles M-270 |
|
Bead form |
Porous |
Porous; sold iron core |
Solid |
|
Ligand |
GFP VHH |
GFP VHH |
GFP VHH |
|
Affinity to GFP |
1 pM |
1 pM |
1 pM |
|
GFP-tagged protein size* |
Small to large size |
Small to large size |
Small to very large size; all sizes |
|
Color |
White |
Black |
Brown |
|
Medium particle size |
~90 µm |
~40 µm |
2.8 µm |
|
Binding capacity |
12 µg/ 10 µL |
8 µg/ 10 µL |
1 μg/ 10 μL |
|
Background |
Very low |
Low |
Low |
|
Magnetic separation & automation |
no |
Yes |
Yes |
|
May be centrifuged up to |
2,500 x g |
800 x g |
8,000 x g |
* Does depend on protein size and shape, protein multimers, complexes and interaction partners
Please contact us for more details.
Find out for yourself and test the GFP-Trap:
ChromoTek and GFP-Trap are registered trademarks of ChromoTek GmbH, and Nanobody is a registered trademark of Ablynx, a Sanofi company.
