GFP-Trap® is a high quality GFP-binding protein based on a single domain antibody derived from Camelids. It is characterized by a small barrel shaped structure (13 KDa, 2.5nm X 4.5 nm) and a very high stability (stable up to 70°C, functional within 2M NaCl or 0.5% SDS). From detailed in vitro binding analysis, it has been determined that one molecule of GFP-Trap® binds one molecule of GFP in a stable, stoichiometric complex with a dissociation constant (Kd) of 0.59 nM. After coupling to monovalent matrices (e.g. agarose beads or magnetic particles), GFP-Trap® can be a robust tool for:
With much greater stability, speciﬁcity, and afﬁnity, GFP-Trap®, the recent addition to antibodies for immunoprecipitation, should make GFP the most suitable tag for immunoprecipitation assays. Direct comparison of the GFP-Trap® with conventional antibodies is shown below. Besides the original avGFP from jellyﬁsh, GFP-Trap® can also bind to GFPS65T and eGFP versions; as well as YFP and eYFP. It recognizes and binds a three dimensional epitope at the GFP beta barrel structure. GFP-Trap® does not recognize unfolded or denatured GFP (e.g. on Western blots).
|Immunoprecipitations (IP) of GFP from protein extracts of GFP-producing human cells. 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.||
Left (IP): Pulldown of 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.
RFP-binding protein based on a single domain antibody derived from camelid single domain antibodies.
The RFP-Trap pulls down all major variants
A VHH domain binding protein derived from camelid heavy chain-only antibodies is coupled to a strong fluorescent dye to both stabilize GFP and enhance its fluorescence signals.
GFP is a powerful tool to study protein localization and dynamics in living cells. However, the photo stability and the quantum efficiency of GFP is not sufficient for Super-Resolution Microscopy (e.g. 3D-SIM or STED) of fixed samples from cells expressing GFP-fusion proteins to visualize specific structures. Furthermore, many cell biological methods such as HCl treatment for BrdU-detection, the EdU-Click-iT™ treatment or heat denaturation for FISH lead to disruption of the GFP signal.
GFP-Booster _Atto488 is a specific GFP-binding protein coupled to the fluorescent dye ATTO 488 (from ATTO-TEC) for reactivation, boosting and stabilization of GFP.
ATTO 488: Fluorescence is excited most efficiently in the range 480 - 510 nm. For instance a 488 nm laser is very suitable for excitation (λabs= 501 nm). Fluorescence is emitted most efficiently in the range 520 - 560 nm (λfl= 523 nm)
GFP-Trap® immobilized in wells to test your GFP fusion proteins for peptide, protein, DNA or RNA binding.
GFP-multiTrap® is available in black 96-well plate format with clear bottom for colorimetric, chemiluminescence and fluorescence detection methods. It is useful for fast and easy capture of GFP-tagged proteins and complexes in a high throughput format to analyse protein interactions (incl. DNA, RNA or peptide binding). The advantages of GFP-multiTrap are:
Fusion of proteins to Glutathione S-Transferase (GST) of Schistosoma japonicum is a common technique to increase solubility and expression level of a protein. Furthermore, GST- tagged fusion proteins are often used in protein-protein interaction studies. Both approaches require highly specific tools to isolate and detect GST fusion proteins. ChromoTek now offers GST-Trap and a rat monoclonal GST antibody for powerful biochemical and Western Blot studies.
GST-Trap is a high quality GST-binding protein based on a single domain antibody derived from Camelid sp. It is characterized by a small barrel shaped structure (13 KDa, 2.5 nm X 4.5 nm) and a very high stability (stable up to 70°C, functional within 2M NaCl or 0.5% SDS).
By now it is clear that Dnmt1, the major eucaryotic DNA methyltransferase, faithfully maintains genome-wide methylation patterns and plays an essential role in the epigenetic network controlling gene expression and genome stability during development. However, the molecular mechanisms that ultimately control DNA methylation still remain elusive. Use the Dnmt1-Trap for very fast, efficient and specific pulldown of endogenous Dnmt1 from cell extracts. CoIP interacting factors of Dnmt1 and identify and characterize them e.g. through mass spec.