GFP-Trap Affinity Reagents

Novel Antibody Fragment that binds GFP Fusion Proteins

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Analyze GFP fusion proteins – in a tube
Chimeric proteins containing naturally-evolved fluorescent proteins are popular tools to study protein localization and dynamics using microscopy. These constructs usually fuse the entirety–or at least a functional domain–of a target protein to one of the multitude of fluorescent proteins. Those originally derived from the jellyfish A. victoria are referred to as green fluorescent protein, or GFP. Cellular analysis of the GFP-fusion protein construct is easily accomplished with a host of available live cell imaging solutions. To get the full picture, though, such data have to be combined with additional biochemical information for the respective target proteins, typically fused to a different “tag” domain that allows for purification. These additional in vitro analyses can be used to confirm functionality of the “tagged” fusion construct, as well as to pull out multi-protein complexes that may form in the cellular milieu. So far, the lack of specific, reliable, and efficient reagents has limited the combination of cell biology results with direct biochemical analysis. Until now, that is.

GFP-Traps utilize super-high affinity Camelidae antibody fragments coupled to agarose beads or magnetic agarose beads. These “Nanobody-Traps” are perfect for immuno-precipitation, immuno-purification, and immuno-pull down experiments with up to 10-fold better purity and yield than that of classic mouse monoclonal antibodies. Compatible with a variety of source materials, Nanobody-Traps may be used with mammalian cells, tissues & organs, bacteria, yeast, and even plants. These reagents allow your GFP-fusions to be perfect candidates for immunoprecipitations, Co-IP, mass spectroscopy, enzyme activity measurements, and ChIP analysis.

GFP-Trap Affinity Reagents are perfect for fast, clean, and efficient one-step isolation of these fluorescent fusion proteins and their interacting factors. With eight years of production experience and over 1000 publications attesting to their broad use and effectiveness, we invite you to try this unique reagent for free. They’ve become a staple of cell biology research throughout Europe and North America.

GFP-Trap has near-perfect affinity and a phenomenal Kd of 1pM over a range of fusion protein concentrations!


Nanobody-traps allow GFP to be used as affinity tags
GFP proteins are not traditionally considered as affinity tags to purify fusion proteins. Nanobody-Traps have changed this notion. To the left is a comparison of GFP-Trap with conventional monoclonal and polyclonal antibodies. Immunoprecipitations (IP) of GFP from protein extracts of GFP-producing human cells was performed. Input (I), non-bound (FT), and bound (B) fractions were separated by SDS-PAGE followed by Coomassie staining and Western blotting. (hc) heavy chain, (lc) light chain of conventional antibodies. The GFP-Trap protocol took 1/10th the time of traditional antibody-based immunoprecipitation, yielded better purity, and resulted in 10-fold higher recovery as indicated by Western analysis.


Protein complexes can’t hide from Nanobody-Traps
To the right are results from using GFP-Trap bound to magnetic particles (GFP-Trap M) and agarose beads (GFP-Trap A). Comparison of GFP-Trap A and GFP-Trap M was first performed with a simple, 1-step pull-down of native GFP with GFP-Trap A and GFP-Trap M from 293T cell extracts. Input (I) and bound (B) fractions were separated by SDS-PAGE followed by Coomassie staining. As expected, the bound fractions containing GFP are extraordinarily pure. The same 1-step purification procedure was then applied to GFP fused to the protein PCNA in a co-immunoprecipitation experiment. This time, the chimera and any associated proteins were very efficiently purified with both GFP-Trap A and GFP-Trap M from 293T cell extracts. As can be seen from the gel, the additional bands represent potential binding partners for the GFP-PCNA.


What’s your lab’s favorite flavor?
Nanobody-Traps are high quality binding proteins coupled to a monovalent matrix (agarose beads, magnetic particles) for biochemical analysis of many fusion proteins and their interacting partners. The different formats allow you to perform a multitude of experiments. The most popular forms are Nanobody-Traps linked to agarose beads. Agarose beads provide the largest binding capacity and lowest background, while still being easy to work with. If automation is important to your lab, then magnetic agarose particles are a great choice. The purified protein complexes are compatible with down-stream analyses such as mass spectroscopy.


Camelidae single-domain antibodies are like IgGs on steroids

The family of animals known as Camelidae (camels, llamas, and alpacas) produce functional antibodies devoid of light chains, so-called "heavy chain" antibodies. These heavy chain antibodies recognize and bind their antigens via a single variable domain. When cleaved from their carboxy tail, these barrel-shaped structures (2x3 nm) are extraordinarily small, naturally-occurring, and intact antigen binding fragments (MW of 13 kDa). These fragments, called Nanobodies, are characterized by high specificity, affinities in the low nanomolar range, and dissociation constants in the sub-nanomolar range (typically 10- to 100-fold better than mouse IgGs). The compact size of Nanobodies makes them extremely stable at temperatures up to 70°C, and functional even in 2M NaCl or 0.5% SDS. These small and powerful antibody fragments can be used in a variety of unique applications. They will open up your research possibilities.



GFP-Traps Coupled to Agarose

GFP-Traps Coupled to Magnetic Agarose Beads

GFP-Traps Non-Coupled

Binding Controls

Protein Controls

Spin Columns

SPECIFICATIONS
Configuration:

GFP-Trap A

Specific Camelidae antibody linked to agarose bead
Part Numbers
GTA010, GTA020, GTA100, GTA200, GTA400, GTAK020

GFP-Trap MA

Specific Camelidae antibody linked to magnetic agarose particle
Part Numbers
GTMA010, GTMA020, GTMA100, GTMA200, GTA400, GTMAK020

GFP-Trap

Uncoupled and purified Camelidae antibody
Part Numbers
GT250
Specificity:

GFP-Traps

GFP, eGFP, wtGFP, GFP S65T, TagGFP, AcGFP, eYFP, YFP, Venus, Citrine, eCitrine, CFP

(does not recognize TurboGFP or all RFPs)

Particle Size:

GFP-Trap A

~90 µm

GFP-Trap MA

~40 µm

GFP-Trap

No particle coupled
Storage Buffer:

GFP-Trap A

20% EtOH

GFP-Trap MA

20% EtOH

GFP-Trap

1x PBS; Preservative: 0.01% Sodium Azide
Storage and Stability:

GFP-Trap A

store at 4°C; stable for one year. Do not freeze.

GFP-Trap MA

store at 4°C; stable for one year. Do not freeze.

GFP-Trap

store at 4°C; stable for one year. Do not freeze.