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Watch Episode 166 When’s the last time you argued in the lab about injecting GTPγS? Four hundred plates of assays, concentration-response curves, and mutagenesis screens are standard. _____________ Keyword Cloud: GPCR scientist network, GPCR training program, mentorship in science, GTPγS assay, lab culture

discovery, mu-opioid receptor, positive allosteric modulators, GPCR training program, GPCR online course, GTPγS assay, chronic pain

Thus we propose the use of fluorescent probes in GPCR screening assays due to their numerous advantages The advantages of Fluorescent Probes in GPCR assays over other methods Fluorescent ligands are made by receptors, tracking of their internalization and are also safer alternatives to radioligand binding assays The traditional binding assays for GPCRs use radioligands, whose limitations, especially safety concerns common starting point for employing fluorescent probes in GPCR assays.

Radioligands have been used to study GPCRs for decades, but with the advances in the fluorescence field, assays They are ligands labeled with radioactive isotopes which can be used in binding assays to quantify other Fluorescent Probes: Differences in Binding Assays Fluorescent ligands are a type of fluorescent probes that offer an alternative to radioligands in binding assays. GPCR-radioligand binding assays.

Imagine running a calcium assay and discovering your compound shows only weak activity. Kenakin dives deep into a widely used, often misunderstood tool in early drug discovery: the calcium assay Revered for its convenience, the FLIPR assay provides rapid insights into receptor activity. with strategic clarity. 📍 Foundational Level | Calcium Assays 📚 Part of Terry Kenakin’s Pharmacology Unlock "Calcium Assays" now

Enhance your GPCR research with deeper assay insights for more effective results. Terry's Corner – Assay Volume Control in GPCR Drug Discovery This week in Terry’s Corner, you’ll learn But behind every “new” assay is a decade of design, failure, and rethinking.

But what if the assay system itself  could be leveraged as a powerful experimental variable? Assay volume control does just that. The real question: are you letting your assay system dictate the wrong story? Clarify whether observed effects reflect true pharmacology or assay artifacts . Unlock Assay Volume Control Only in Terry’s Corner 🎬 Watch Course Trailers Not sure yet?

Overcoming them requires careful assay design and strategic use of the right fluorescent probes. For GPCR assay developers, HCS supports:  Quantitative visualization of receptor internalization and trafficking  • Live-cell kinetic measurements  unavailable to endpoint assays  Multiplexed assessment Why Traditional Radioligand Methods Fall Short for Modern Screening Needs Radioligand binding assays the gold standard for image-based GPCR assays.

is to obtain a fluorescent ligand suitable for the Tag-lite® binding assays. (Ki) and saturation Tag-lite® binding assay (Kd).   1  Competition radioligand binding assay.  2  Saturation assay by Tag-lite® binding assay.  3  Displacement of specific [3H]-CP55940 binding in human HEK-CB1 assay and Kd= 42nM in Tag-lite®). Saturation assays using CELT-335.

Homogeneous Time-Resolved Fluorescence Assays: Overcoming Common Challenges Balancing sensitivity and This technology enhances sensitivity and assay specificity . Saturation assays using CELT-335. A Robust and Efficient FRET-Based Assay for Cannabinoid Receptor Ligands Discovery. A Robust and Efficient FRET-Based Assay for Cannabinoid Receptor Ligands Discovery.

August 2022 In vitro assays for the functional characterization of (psychedelic) substances at the serotonin More specifically, this review covers assays to monitor the canonical G protein signaling pathway (e.g measuring G protein recruitment/activation, inositol phosphate accumulation, or Ca2+ mobilization), assays recruitment or signaling, and assays to monitor receptor conformational changes. mechanism and linked to its specific execution, as these may heavily impact the assay outcome."

Thus, developing assays for commercially available probes such as CELT-419 would facilitate research Fluorescence anisotropy (FA) assay has been used to study the ligand binding kinetics of many GPCRs.[ assays, the radioligand binding method was used.  The estimated Z’ of the assay is 0.71, which is sufficient for HTS standards. Figure 3.   provide more information and alternatives for binding assays.

Eric Trinquet, a veteran innovator behind functional GPCR assays like HTRF and IP-One, believes rigid But those failures are where new paths emerge, often leading to transformative tools like the IP1 assay Eric Trinquet Built to Fail, Built to Win: Inside the IP1 Assay Origin Story The IP-One assay didn’t Instead, they focused on equilibrium-based assays and zeroed in on IP1 accumulation—pioneering a clean “We did a full dose-response and saw antagonism—all in one plate-based assay.

Every GPCR assay that makes it to market carries years of failures, late-night ideas, risky bets, and Functional assays for GPCRs—especially Gq-coupled receptors—were notoriously messy. Calcium flux? Trinquet’s team asked a bolder question: could they design equilibrium-based assays for pathways no one Built to Fail, Built to Win: The IP One Gamble After the success of their cAMP assay, Trinquet’s team Months were spent debating assay design. IP1 isn’t naturally abundant or easy to detect.

kinetics ✅ A framework to classify antagonists and rank compounds by offset rate, using rapid calcium assays The Hemi-Equilibrium Problem Calcium assays look simple. Flip it around, and you’ve got a shortcut: use depression of max in calcium assays to rapidly rank offset The reality is dynamic—ligands arrive, depart, and interact in ways that standard assays often miss. suggestions Practical insights  distilled from decades of pharmacology experience Whether you’re validating assays

Does Your Assay Agree With the Literature? Your experiment gives a pKB of 8.0. Is that consistent, or is your assay biased? to: Confirm whether your mean truly overlaps with accepted values Verify if binding and functional assays interchangeable with existing ones Instead of vague comparisons, you’ll have statistical clarity on whether your assay suggestions Practical insights  distilled from decades of pharmacology experience Whether you’re validating assays

developed and validated a label-free, liquid chromatography-mass spectrometry (LC-MS) based cell binding assay This assay was applied to various target classes, with particular emphasis on those for which protein-based binding assay can be difficult to achieve. In another example, this assay was applied to an ion channel target with its agonists, of which the determined binding affinity was consistent with functional assays.

August 2022 Luciferase-based GloSensor™ cAMP assay: Temperature optimization and application to cell-based The GloSensor™ cAMP assay enables real-time monitoring of signaling downstream of many GPCRs. However, it is crucial to optimize assay conditions such as temperature. Here, we describe the temperature sensitivity and reversibility of the GloSensor™ cAMP assay, and which Nevertheless, the GloSensor™ cAMP assay can be applied to analyze signaling by a wide range of GPCRs

Watch Episode 176 The first time Michelle ran a cyclic AMP assay, she did it with a single-channel pipette Assay samples were layered on trays of ice. These weren’t quaint inconveniences. We were doing assays on ice, pipetting one sample at a time. Those early cyclic AMP assays weren’t just cold and slow — they were part of a bigger puzzle. Mini Timeline: Manual assay years — technical rigor as foundation Technology boom — scaling curiosity

. • Assay Development Gets Real : Fluorescent tools and real-world biology don’t always match. Whether you’re designing the next assay, scouting a new therapeutic angle, or exploring career pivots

Traditional assays—particularly membrane-based radioligand binding—often provide high-throughput measurements study, 16 synthetic cannabinoid receptor agonists (SCRAs) were evaluated using a CB2 live-cell HCS assay Traditional membrane-based assays isolate receptors from this environment, which simplifies quantification Visual displacement provides an additional confidence layer that traditional homogeneous binding assays or GPCR-mediated analgesia, these findings reinforce the value of physiologically relevant binding assays

October 2022 "We describe production of the human A2A adenosine receptor (A2AAR), a class A G protein-coupled receptor (GPCR) for 19F-NMR and single-molecule fluorescence (SMF) spectroscopy. We explain in detail steps shared between the two sample preparation strategies, including expression and isolation of A2AAR and assembly of A2AAR in lipid nanodiscs and procedures for incorporation of either 19F-NMR or fluorescence probes. Protocols for SMF experiments include sample setup, data acquisition, data processing, and error analysis. For complete details on the use and execution of this protocol, please refer to Wei et al. (2022) and Sušac et al. (2018)." Read more at the source #DrGPCR #GPCR #IndustryNews

pHSense™ reagents enable real-time GPCR internalization detection A breakthrough GPCR internalization assay Developed to address long-standing challenges in GPCR internalization assays , pHSense™ reagents  combine Built on more than two decade A GPCR Internalization Tool Designed for Real Research Needs s of GPCR assay assays β-arrestin recruitment assays Downstream readouts  – phospho-ERK, AKT, CREB, MEK and other phoshoproteins , where he shares the story behind pHSense , including its application in real-time internalization assays

However, these conventional assays often provide limited information on intermediate signaling events (e.g G protein recruitment assay [6], mini-G recruitment assay [7], GRK and β-arrestin recruitment assays [8]), G protein activation (e.g TRUPATH assay [9]), receptor trafficking ( e.g FYVE assay [10]),  β-arrestin  YEN assay [13]), among others. Zhou, Y., et al., Multiple GPCR Functional Assays Based on Resonance Energy Transfer Sensors. 

To efficiently run these screening campaigns high throughput screening assays are used. How Fluorescence Polarization Assays Work: Principles and Applications in GPCR Research FP assays work FP assays are convenient and easy to manipulate. where they used CELT-419 for D3 dopamine receptor binding assays in baculoviruses. 3.       The future of fluorescence polarization assays is closely tied to fluorescent ligands.

During that project we tested its validity as a fluorescent probe for Tag-lite® assays, where we used a set of 7 cannabinoid ligands (both natural and synthetic) to validate and optimize the assay. It bears a hydrophilic fluorophore compatible with the terbium donor in TR-FRET assays. Figure 2.Assay Principle: TR-FRET assay using CELT-335, a dual (CB1/CB2) fluorescent ligand that serves Comparison of binding affi nities using radioligand binding or TR-FRET assays.

It was simply a clean, scalable assay—and a wave of new possibilities. Instead of creating another black-box assay, they designed pHSense around rare-earth europium probes. This was the foundation of a new set of assay tools. Whether in industry or academia, whether overexpressing or not, you can adapt the assay to your system Build or adopt assays that can evolve with your questions—like pHSense.” What’s Next?

programs don’t fail because of weak molecules—they fail because biology behaves differently than the assay His work shows why chemical design can outperform antibodies and how rigorous assay validation bridges But HCS only works when assays are built with rigor and powered by the right fluorescent ligands. What You’ll Learn Why traditional radioligand assays miss critical spatial and kinetic signals Five phases a robust, reproducible HCS pipeline How fluorescent ligands strengthen specificity, relevance, and assay

Docking scores were meaningless without assays. For receptor assays and the biological interpretation of ligands, he relies on a network of collaborators Chemistry fuels assays. Assay data flows back into models. A pharmacologist who understands docking, or a modeler who knows assay constraints, will collaborate , and assay data cycling back to refine predictions.

“We don’t just deliver compounds, we solve assay problems.” — Dr. Democratize access to high-performance chemical probes and make assay development faster, cheaper, and workflow on the company page . _______________ Keyword Cloud:   GPCR data platform , fluorescent ligands , assay