CINXE.COM

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block-system clearfix"> <div class="col-xs-12 col-sm-8 col-md-9 col-lg-9 col-xl-9 et_pharm_browser_wrapper"> <h1>TR-FRET and FP Assay Reagents</h1> <p>TR-FRET (Time-Resolved Fluorescence Resonance Energy Transfer) and FP (Fluorescence Polarization) assays are widely used in biology and drug discovery to study target engagement. Tocris provides a range of fluorescent probes, including CoraFluor™ terbium cryptate FRET donors and fluorescent E3 ligase ligands, suitable for exploring ligand binding in TR-FRET and FP assays.</p> <div class="tabbed_content"> <div class="tab_menu"> <div class="tab tab_1 tab_open" data-tab="1" data-tabname="products" id="pharm_tabs_products"> <span>Products</span></div> <div class="tab tab_2" data-tab="2" data-tabname="background" id="pharm_tabs_background"><span>Background</span> </div> <div class="tab tab_4" data-tab="4" data-tabname="literature" id="pharm_tabs_literature"><span>Literature (2)</span> </div> </div> <div class="tab_content tab_1 tab_open" data-tab="1"> <div class="pharm_products"><table class="pharm_prod_table table table-condensed"> <tr> <th>Cat. No.</th> <th>Product Name / Activity</th> </tr> <tr> <td>7857</td> <td><a href="/products/bdy-fl-lenalidomide_7857" title="View price & availability information for BDY FL Lenalidomide">BDY FL Lenalidomide</a></td> </tr> <tr class="pharm_prod_table_action"> <td></td><td>Fluorescent cereblon ligand</td> </tr> <tr> <td>7985</td> <td><a href="/products/bdy-fl-staurosporine_7985" title="View price & availability information for BDY FL Staurosporine">BDY FL Staurosporine</a></td> </tr> <tr class="pharm_prod_table_action"> <td></td><td>Fluorescent probe targeting kinases; kinase tracer for TR-FRET assays</td> </tr> <tr> <td>7633</td> <td><a href="/products/bdy-fl-thalidomide_7633" title="View price & availability information for BDY FL Thalidomide">BDY FL Thalidomide</a></td> </tr> <tr class="pharm_prod_table_action"> <td></td><td>High affinity fluorescent probe for TR-FRET cereblon binding assay</td> </tr> <tr> <td>7483</td> <td><a href="/products/bdy-fl-vh032_7483" title="View price & availability information for BDY FL VH032">BDY FL VH032</a></td> </tr> <tr class="pharm_prod_table_action"> <td></td><td>High-affinity VHL fluorescent probe for TR-FRET and FP assays</td> </tr> <tr> <td>7952</td> <td><a href="/products/celt-133_7952" title="View price & availability information for CELT-133">CELT-133</a></td> </tr> <tr class="pharm_prod_table_action"> <td></td><td>Selective hα<sub>1A</sub> adrenergic receptor fluorescent antagonist; suitable for use in TR-FRET</td> </tr> <tr> <td>7953</td> <td><a href="/products/celt-211_7953" title="View price & availability information for CELT-211">CELT-211</a></td> </tr> <tr class="pharm_prod_table_action"> <td></td><td>Fluorescent serotonin 5HT<sub>2B</sub> receptor ligand for HTS; suitable for use in TR-FRET</td> </tr> <tr> <td>7954</td> <td><a href="/products/celt-327_7954" title="View price & availability information for CELT-327">CELT-327</a></td> </tr> <tr class="pharm_prod_table_action"> <td></td><td>Fluorescent hA<sub>2B</sub>/A<sub>3</sub> adenosine receptor antagonist; suitable for use in TR-FRET</td> </tr> <tr> <td>7955</td> <td><a href="/products/celt-426_7955" title="View price & availability information for CELT-426">CELT-426</a></td> </tr> <tr class="pharm_prod_table_action"> <td></td><td>Potent, partially selective and fluorescent hD<sub>2</sub> dopamine receptor antagonist; suitable for use in TR-FRET</td> </tr> <tr> <td>7920</td> <td><a href="/products/corafluor-1-amine-reactive_7920" title="View price & availability information for CoraFluor™ 1, amine reactive">CoraFluor™ 1, amine reactive</a></td> </tr> <tr class="pharm_prod_table_action"> <td></td><td>Terbium cryptate FRET donor for TR-FRET assay development; amine reactive for conjugation</td> </tr> <tr> <td>8117</td> <td><a href="/products/corafluor-1-thiol-reactive_8117" title="View price & availability information for CoraFluor™ 1, thiol reactive">CoraFluor™ 1, thiol reactive</a> <span class="prod-new-logo" title="New: CoraFluor™ 1, thiol reactive">New</span></td> </tr> <tr class="pharm_prod_table_action"> <td></td><td>Terbium cryptate FRET donor for TR-FRET assay development, cysteine reactive for conjugation</td> </tr> <tr> <td>7950</td> <td><a href="/products/corafluor-2-amine-reactive_7950" title="View price & availability information for CoraFluor™ 2, amine reactive">CoraFluor™ 2, amine reactive</a></td> </tr> <tr class="pharm_prod_table_action"> <td></td><td>Terbium cryptate FRET donor for TR-FRET assay development; amine reactive for conjugation</td> </tr> <tr> <td>7287</td> <td><a href="/products/fam-deala-hyp-yipd_7287" title="View price & availability information for FAM-DEALA-Hyp-YIPD">FAM-DEALA-Hyp-YIPD</a></td> </tr> <tr class="pharm_prod_table_action"> <td></td><td>Fluorescent HIF-1α peptide; can be used to assess VHL binding in FP assays</td> </tr> <tr> <td>6574</td> <td><a href="/products/fluorescein-nad-plus_6574" title="View price & availability information for Fluorescein-NAD+">Fluorescein-NAD+</a></td> </tr> <tr class="pharm_prod_table_action"> <td></td><td>Fluorescent NAD<sup>+</sup>; substrate for ADP-ribosylation for use in PARP assays; wavelengths are compatible with use as an acceptor dye in TR-FRET assays</td> </tr> <tr> <td>7722</td> <td><a href="/products/jq1-fitc_7722" title="View price & availability information for JQ1-FITC">JQ1-FITC</a></td> </tr> <tr class="pharm_prod_table_action"> <td></td><td>Fluorescent BET bromodomain probe; suitable for use in TR-FRET</td> </tr> <tr> <td>4577</td> <td><a href="/products/ldv-fitc_4577" title="View price & availability information for LDV FITC">LDV FITC</a></td> </tr> <tr class="pharm_prod_table_action"> <td></td><td>Flourescent α<sub>4</sub>β<sub>1</sub> integrin ligand; wavelengths are compatible with use as an acceptor dye in TR-FRET assays</td> </tr> <tr> <td>8126</td> <td><a href="/products/mpro-tracer_8126" title="View price & availability information for Mpro Tracer">Mpro Tracer</a> <span class="prod-new-logo" title="New: Mpro Tracer">New</span></td> </tr> <tr class="pharm_prod_table_action"> <td></td><td>TR-FRET acceptor; SARS-CoV-2 main protease fluorescent ligand</td> </tr> <tr> <td>6461</td> <td><a href="/products/parpi-fl_6461" title="View price & availability information for PARPi-FL">PARPi-FL</a></td> </tr> <tr class="pharm_prod_table_action"> <td></td><td>Fluorescent potent PARP inhibitor; wavelength compatible with use as an acceptor dye in TR-FRET assays</td> </tr> <tr> <td>7970</td> <td><a href="/products/saha-fitc_7970" title="View price & availability information for SAHA-FITC">SAHA-FITC</a> <span class="prod-new-logo" title="New: SAHA-FITC">New</span></td> </tr> <tr class="pharm_prod_table_action"> <td></td><td>Fluorescent HDAC1/2 probe; suitable for use in TR-FRET and FP assays</td> </tr> <tr> <td>7288</td> <td><a href="/products/thalidomide-cyanine-5_7288" title="View price & availability information for Thalidomide-Cyanine 5">Thalidomide-Cyanine 5</a></td> </tr> <tr class="pharm_prod_table_action"> <td></td><td>High affinity cereblon fluorescent probe for use in TR-FRET</td> </tr> <tr> <td>2540</td> <td><a href="/products/tocrifluor-t1117_2540" title="View price & availability information for Tocrifluor T1117">Tocrifluor T1117</a></td> </tr> <tr class="pharm_prod_table_action"> <td></td><td>Fluorescent cannabinoid CB<sub>1</sub> receptor ligand; wavelength compatible with use as an acceptor dye in TR-FRET assays</td> </tr> <tr> <td>8069</td> <td><a href="/products/xiap-tracer-mf-smac_8069" title="View price & availability information for XIAP Tracer mF-Smac">XIAP Tracer mF-Smac</a> <span class="prod-new-logo" title="New: XIAP Tracer mF-Smac">New</span></td> </tr> <tr class="pharm_prod_table_action"> <td></td><td>Fluorescein labeled XIAP ligand for TR-FRET assay development</td> </tr></table></div> </div> <div class="tab_content tab_2" data-tab="2"> <h2>What is TR-FRET?</h2> <p>TR-FRET (Time-Resolved Fluorescence Resonance Energy Transfer) is a highly sensitive and widely used assay technology in the field of drug discovery and biological research to study protein-protein interactions, enzyme kinetics, and receptor-ligand binding. This technology works by measuring the energy transfer between two fluorescent molecules - a donor (such as CoraFluor™) and an acceptor. When the donor molecule is excited by a light source, typically 340 nm, energy is transferred from the donor to the acceptor through a phenomenon called Förster resonance energy transfer (FRET) if the two molecules are in proximity (typically 5-10 nm) to each other (Figure 1). TR-FRET assays generally utilize lanthanides as donors, which have a large Stokes shift and millisecond long lifetimes, enabling time-delayed detection and eliminating background noise derived from scattered excitation light and autofluorescence from media components and proteins. </p> <p>Our TR-FRET toolbox offers a wide range of highly versatile probes to generate biochemical assays.</p> <img src='https://resources.tocris.com/images/targets/figure-1-tr-fret-assay-principle.png' alt='Schematic outlining the TR-FRET assay principle as applied to induced proximity' style='max-width:100%; margin:20px 0 10px 0'> <p><strong>Figure 1:</strong> Schematic outlining the TR-FRET assay principle as applied to induced proximity. </p> <h2>Featured Product: CoraFluor™ Probes</h2> <p><a href='/products/corafluor-1-amine-reactive_7920' title='View product'>CoraFluor™ 1</a> (Cat. No. 7920) and <a href='/products/corafluor-2-amine-reactive_7950' title='View product'>CoraFluor™ 2</a> (Cat. No. 7950) are terbium-based TR-FRET donors that emit wavelengths compatible with commonly used fluorescent acceptor dyes such as FAM or <a href='/products/fitc_5440' title='View product'>FITC</a> (Cat. No. 5440), <a href='/product-type/bdy-dyes' title='Browse BODIPY dyes'>BODIPY<sup>®</sup></a> (BDY), <a href='/product-type/janelia-fluor-dyes' title='Find Janelia Fluor dyes'>Janelia Fluor<sup>®</a></sup> dyes, TMR, and <a href='/products/cyanine-5-se_5436' title='View product'>Cyanine 5, SE</a> (Cat. No. 5436), making it easy to incorporate into ongoing screening cascades (Figure 2). Compared to existing TR-FRET donors, the CoraFluor fluorescence is brighter and more stable in biological media, enhancing sensitivity and data generation from biochemical assays. CoraFluor™ 1 exhibits excitation upon exposure to a 337 nm UV laser, whereas CoraFluor™ 2 displays a red-shifted excitation wavelength, enhancing excitation efficiency at 365 nm and 405 nm. This attribute of CoraFluor™ 2 widens the range of analytical instruments suitable for these assays.</p> <p>CoraFluor reagents can be conjugated directly to your antibody or protein of interest (see <a href='/resources/protocols/fluorescent-dye-conjugation/conjugation-protocol-amine-reactive-corafluor' title='See protocol'>protocol</a>). Alternatively, custom services for the conjugation of CoraFluor reagents to an antibody of your choice are available from <a href='https://www.rndsystems.com/services/custom-antibody-services' target='_blank' title='Learn more about Custom Conjugation services from R&D Systems'>R&D Systems</a>. </p> <img src='https://resources.tocris.com/images/targets/figure-2-corafluor-emission-spectra.png' alt='CoraFluor emission spectral overlap with common acceptor dyes' style='max-width:100%; margin:20px 0 10px 0' loading='lazy'> <p><strong>Figure 2:</strong> CoraFluor emission spectral overlap with common acceptor dyes.</p> <h2>What is TR-FRET Used For?</h2> <p>TR-FRET has been extensively used to identify novel drug targets, screen compound libraries, and optimize lead compounds in various therapeutic areas, including oncology, cardiovascular disease, infectious disease, and neurodegenerative disorders. TR-FRET assays are particularly useful in measuring the binding affinity of small molecules, such as inhibitors and Degraders, to protein targets in a multi-well plate format, allowing for efficient high-throughput screening of target engagement (Figure 3).</p> <img src='https://resources.tocris.com/images/targets/figure-3-corafluor-tr-fret-target-engagement-assay.png' alt='CoraFluor TR-FRET target engagement assay' style='max-width:100%; margin:20px 0 10px 0' loading='lazy'> <p><strong>Figure 3:</strong> CoraFluor TR-FRET target engagement assay.</p> <h2>TR-FRET in Degrader Development</h2> <p>TR-FRET ternary complex assays are valuable in the development of small molecule protein <a href='/product-type/protein-degraders' title='Browse our range of protein Degrader molecules'>Degraders</a> as they provide a quantitative measure of the interactions between the Degrader, the target protein, and the <a href='/pharmacology/ubiquitin-e3-ligases' title='Browse products for ubiquitin E3 ligases'>ubiquitin E3 ligase</a> (Figure 3). This information is critical for achieving effective <a href='/product-type/targeted-protein-degradation' title='Find products for Targeted Protein Degradation research'>targeted protein degradation</a> by characterizing the binding affinity of the Degrader to the target protein, the recruitment of ubiquitin E3 ligase to the target protein, and the formation of a cooperative ternary complex necessary for ubiquitination and subsequent degradation. This allows for optimization of Degrader molecules to improve their potency and selectivity. </p> <img src='https://resources.tocris.com/images/targets/figure-4-corafluor-tr-fret-ternary-complex-assay.png' alt='CoraFluor TR-FRET ternary complex assay' style='max-width:100%; margin:20px 0 10px 0' loading='lazy'> <p><strong>Figure 4:</strong> CoraFluor TR-FRET ternary complex assay.</p> <p>It is essential to quantify protein levels accurately. TR-FRET assays present a practical approach for the precise determination of target protein levels in cellular lysates after treatment with a Degrader, leveraging the advantages of a high-throughput format on a fluorescence microplate reader. This technique offers invaluable insights into the time- and dose-dependent kinetics of degradation for a panel of candidate Degraders. Furthermore, it enables calculation of important parameters, including the DC<sub>50</sub> (the concentration at which target protein degradation is half-maximal) and D<sub>max</sub> (the maximal achievable degradation level) (Figure 5). </p> <img src='https://resources.tocris.com/images/targets/figure-5-corafluor-tr-fret-protein-degradation-quantification-assay.png' alt='CoraFluor TR-FRET protein degradation quantification assay' style='max-width:100%; margin:20px 0 10px 0' loading='lazy'> <p><strong>Figure 5:</strong> CoraFluor TR-FRET protein degradation quantification assay.</p> <h2>What are FP Assays Used For?</h2> <p>Fluorescence polarization (FP) assays are also widely used in biology to study molecular interactions, such as protein-protein, protein-DNA, and protein-ligand binding, as well as for monitoring the progress of enzyme reactions. The technique is based on the observation that the effect of polarized light applied to a fluorescent probe is dependent on multiple factors, such as molecular conformation, orientation, and size. FP assays measure the change in orientation of a molecule in time, between absorption and emission events, and indicate the level of binding of the fluorescent molecule. The approach provides a rapid, quantitative and versatile analysis of molecular interactions, and can be used to measure affinity constants (K<sub>i</sub>, K<sub>d</sub>) and IC<sub>50</sub> values for small molecules. As such FP assays are frequently used for high-throughput screening in drug discovery.</p> <p><em>CoraFluor™ is a trademark of Bio-Techne Corp.</em></p> <p><em>BODIPY<sup>®</sup> is a registered trademark of Molecular Probes.</em></p> </div> <div class="tab_content tab_3" data-tab="3"> </div> <div class="tab_content tab_4" data-tab="4"> <h3>Literature for TR-FRET and FP Assay Reagents</h3><p>Tocris offers the following scientific literature for TR-FRET and FP Assay Reagents to showcase our products. We invite you to request* your copy today!</p><p class="small">*Please note that Tocris will only send literature to established scientific business / institute addresses.</p><hr/><div class="lit_item"> <div class="lit_content"> <div class="lit_image"> <a href="https://resources.tocris.com/images/literature/product-guides/fullsize/707399.jpg?v=3" data-toggle="lightbox" data-title="Fluorescent Dyes and Probes Research Product Guide" data-footer="<a href='/literature/literature-request/707399' rel='nofollow' title='Request copy of the Fluorescent Dyes and Probes'>Request copy</a> or <a href='/literature/literature-request/p707399' rel='nofollow' title='Request PDF of the Fluorescent Dyes and Probes'>Request PDF</a> today"><img loading="lazy" src="https://resources.tocris.com/images/literature/product-guides/regular/707399.jpg?v=3" alt="Fluorescent Dyes and Probes Research Product Guide" class="img-fluid"></a></div> <h4>Fluorescent Dyes and Probes Research Product Guide</h4> <p>This product guide provides a background to the use of Fluorescent Dyes and Probes, as well as a comprehensive list of our:</p> <ul class='bullet_list'> <li>Fluorescent Dyes, including dyes for flow cytometry</li> <li>Fluorescent Probes and Stains, including our new MitoBrilliant<sup>TM</sup> mitochondria stains</li> <li>Tissue Clearing Kits and Reagents</li> <li>Aptamer-based RNA Imaging Reagents</li> <li>Fluorescent Probes for Imaging Bacteria</li> <li>TSA Vivid<sup>TM</sup> Fluorophore Kits</li> <li>TSA Reagents for Enhancing IHC, ICC & FISH Signals</li> </ul> <ul class="pb_lit_buttons"><li> <a class="btn btn-quinary" href="/literature/literature-request/707399" rel="nofollow" title="Request copy of the Fluorescent Dyes and Probes Research Product Guide">Request copy</a> </li><li> <a class="btn btn-quinary request-pdf" href="/literature/literature-request/p707399" rel="nofollow" title="Request PDF of the Fluorescent Dyes and Probes Research Product Guide">Request PDF<div class="lit_request_pdf"></div></a> </li> <li><a class="btn btn-quinary" href="/literature/product-guides" title="View all Product Guides & Listings">View all Product Guides & Listings</a></li> </ul> </div></div><div class="lit_item"> <div class="lit_content"> <div class="lit_image"> <a href="https://resources.tocris.com/images/literature/product-guides/fullsize/634432.jpg?v=8" data-toggle="lightbox" data-title="TPD and Induced Proximity Research Product Guide" data-footer="<a href='/literature/literature-request/634432' rel='nofollow' title='Request copy of the TPD and Induced Proximity'>Request copy</a> or <a href='/literature/literature-request/p634432' rel='nofollow' title='Request PDF of the TPD and Induced Proximity'>Request PDF</a> today"><img loading="lazy" src="https://resources.tocris.com/images/literature/product-guides/regular/634432.jpg?v=8" alt="TPD and Induced Proximity Research Product Guide" class="img-fluid"></a></div> <h4>TPD and Induced Proximity Research Product Guide<span class="lit-updated-logo" title="Recently updated">Updated</span></h4> <p>This brochure highlights the tools and services available from Bio-Techne to support your Targeted Protein Degradation and Induced Proximity research, including:</p> <ul class='bullet_list'> <li>Active Degraders</li> <li>TAG Degradation Platform</li> <li>Degrader Building Blocks</li> <li>Assays for Protein Degradation</li> <li>Induced Proximity Tools</li> </ul> <ul class="pb_lit_buttons"><li> <a class="btn btn-quinary" href="/literature/literature-request/634432" rel="nofollow" title="Request copy of the TPD and Induced Proximity Research Product Guide">Request copy</a> </li><li> <a class="btn btn-quinary request-pdf" href="/literature/literature-request/p634432" rel="nofollow" title="Request PDF of the TPD and Induced Proximity Research Product Guide">Request PDF<div class="lit_request_pdf"></div></a> </li> <li><a class="btn btn-quinary" href="/literature/product-guides" title="View all Product Guides & Listings">View all Product Guides & Listings</a></li> </ul> </div></div> </div> <div class="tab_content tab_5" data-tab="5"> </div> <div class="tab_content tab_6" data-tab="6"> </div> </div> </div> <div id="sidebar" class="col-xs-12 col-sm-4 col-md-3 col-lg-3 col-xl-3"> <div class="resources_sidebar_container"> <div class="fp_four_box"> <div class="fp_box_wrapper"><h3>Featured New Products</h3> <div class="fp_featured_product"><a href="/products/vu-0542270_8151">8151 | VU 0542270</a><br /> Selective inhibitor of K<sub>ir</sub>6.1 K<sub>ATP</sub> channels</div> <div class="fp_featured_product"><a href="/products/rna-imaging-probe-1c_8813">8813 | RNA Imaging Probe 1c</a><br /> Fluorogenic RNA imaging probe</div> <div class="fp_featured_product"><a href="/products/lsn-3318839_7959">7959 | LSN 3318839</a><br /> Glucagon-like peptide-1 receptor <br />(GLP1-R) positive allosteric modulator</div> <div class="fp_featured_product"><a href="/products/chir-99021-in-solution_8170">8170 | CHIR 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background information on various imaging techniques.</p><a class="btn btn-quinary" href="/literature/literature-request/707399" rel="nofollow" title="Request copy of the Fluorescent Dyes and Probes Research Product Guide">Request copy</a><a class="btn btn-quinary request-pdf" href="/literature/literature-request/p707399" rel="nofollow" title="Request PDF of the Fluorescent Dyes and Probes Research Product Guide">Request PDF<div class="lit_request_pdf"></div></a></div><div class="item"><div class="highlight">Targeted Protein Degradation and Induced Proximity Guide</div> <a href="/literature/product-guides/tpd-and-induced-proximity"><img loading="lazy" src="https://resources.tocris.com/images/literature/product-guides/regular/634432.jpg?v=8" alt="TPD and Induced Proximity Research Product Guide"></a><p class="small">This brochure highlights the products and services available from Bio-Techne to support Targeted Protein Degradation and Induced Proximity research.</p><a class="btn 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