GFP-Trap and RFP-Traps utilize super-high affinity Camelidae antibody fragments coupled to agarose beads, magnetic agarose beads, magnetic particles. These 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, Nano-Traps may be used with mammalian cells, tissues & organs, bacteria, yeast and even plants. These reagents allow your GFP- or RFP-fusions to be perfect candidates for immunoprecipitations, Co-IP, mass spectroscopy, enzyme activity measurements, and ChIP analysis.
GFP-Trap® and RFP-Trap Affinity Reagents are perfect for fast, clean and efficient one-step isolation of these fluorescent fusion proteins and their interacting factors. With over five years of production experience and over 400 publications attesting to their broad use and effectiveness, we invite you to try either of these reagents. They've become a staple of cell biology research in Europe and the UK. The NanoTraps come in several different versions including attached to agaorse or magnetic beads or to the bottom of a plate well, and even as a free complex, providing you the ability to maximize their use.
RFP-Trap to immunoprecipitate red fluorescent proteins
Pulldown of different monomeric red fluorescent proteins mRFP, mCherry and mOrange from cell extracts of human cells. Input (I), non-bound (FT) and bound (B) fractions were separated by SDS-PAGE followed by Coomassie staining and Western Blotting.
Nano-Traps allow GFP and RFP to be used as affinity tags
GFP and RFP proteins are not traditionally considered as affinity tags to purify fusion proteins. Nano-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, with better purity, and resulted in 10-fold higher recovery as indicated by western analysis.
Protein complexes can’t hide from Nano-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 extraordinaly pure. The same 1-step purification procedure was now applied to GFP fused to the protein PCNA in a co-immunoprecipitation type 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 for the gel, the additional bands represent potential binding partners for the GFP-PCNA.
What’s your lab’s favorite flavor?
Nano-Traps are high quality binding proteins coupled to a monovalent matrix (agarose beads, magnetic particles or in 96-well plates) 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 when the Nano-Traps are 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 particles and magnetic agarose particles are a great choice. The magnetic particles are smaller and easier to keep in suspension, while the magnetic agarose particles offer the superior binding kinetics. For high throughput, nothing beats the 96-well plate in which the Nanobodies are bound to the side of the wells. These resins can be used interchangeably for pull-down type experiments such as immunoprecipitations, co-IPs and even ChIP assays. 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, dromedaries, 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 and 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