What GFP-Trap should I use for my immunoprecipitation?


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

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 must 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.

 

 

GFP-Trap beads

 

Agarose

Magnetic Agarose

Magnetic Particles M-270

Matrix

Agarose
(4% cross-linked)

Magnetic agarose
(6% cross linked)

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

 

Find out for yourself and test the GFP-Trap:

Request your free GFP-Trap sample