Reagents for Receptor Binding

G-protein coupled receptors (GPCRs) are crucial transmembrane proteins involved in cellular signal transduction. This technical note outlines a method for preparing GPCR membrane model extracts from stable cell lines, specifically for use with the HTRF GTP Gi binding assay. Get this technical note and discover: Key Highlights such as the Importance of GPCRs and the advantages of using HTRF GTP Gi Binding Assay Detailed Method with Cell Culture Preparation, Cell Lysis, Membrane Preparation and Assay Optimization For research use only. Not for use in diagnostic procedures.

Protein-protein interactions: cover them all with one approach This brochure illustrates the possibilities and versatility of protein-protein interaction studies. It features six relevant examples of various interaction types taken from literature to show you how studies can be handled with the time-resolved fret-based HTRF approach, including virus blockade, receptor/ligand binding, protease activity, and more. Features: Introduction to the stakes of protein-protein interaction research Illustration of 6 published interaction studies involving biologics or small molecules For research use only. Not for use in diagnostic procedures.

Atherosclerosis pathogenesis, cellular actors, and pathways Atherosclerosis is a common condition in which arteries harden and become narrow due to a build-up of fatty material, usually cholesterol, and other substances such as calcium. This can lead to a range of serious health complications, including heart attack or stroke, making the disease an important contributing factor in death and morbidity in developed countries. Recent developments in our understanding of atherosclerosis from a molecular perspective include the discovery of new players in disease pathogenesis. Included in this white paper Atherosclerosis: step-by-step pathogenesis, therapeutic strategies, and recent developments Detailed descriptions and explanations, including a focus on pathways

Taking autophagy regulation research a step further Autophagy regulation is a key molecular process involved in recycling long-lived protein and organelles. Dysregulation of autophagy leads to different pathologies such as cancer, neurodegenerative and infectious diseases. This eBook features: Key facts about autophagy and mitophagy Infographics to apprehend the basics Cutting-edge knowledge

Get a useful overview of today’s immunity knowledge with this booklet Immunity is a collection of complex processes involving multiple strategies and specialized cell types. This booklet provides you with critical information regarding their roles, characteristic and signaling pathways as well as the collaborative behaviors that contribute to immunity. Featured in this guide: Review the fundamentals of immune cell types and mechanisms Learn from a cutting-edge research report Pathways and functional details on over 10 specialized immune cells

Cyclin-dependent kinases (CDKs) 4 & 6 play a key in breast cancer. Cyclin D1-CDK4/6 complexes are critical regulators of the cell cycle transition from the G1 to S phase. To proceed through these phases, a cell must pass a restrictive checkpoint, tightly regulated in this case by the retinoblastoma tumor suppressor protein (Rb). Palbociclib is a small molecule kinase inhibitor that blocks Cyclin D1-CDK4/6 mediated phosphorylation of the Rb protein to prevent E2F driven transcription of genes that commit the cell to DNA replication and cell division. In this application note, you will learn: The effect of palbociclib treatment in two breast cancer cell lines How to monitor the amount of phospho-Rb and Cyclin D1 protein levels with HTRF® ® immunoassays to examine the effect of CDK4/6 inhibition and Cyclin D1 regulation

Over these last few decades there has been a growing trend in drug discovery to use cellular systems and functional assays, in addition to biochemical assays, for the characterization of new potential therapeutics. The ability to study the interaction between a candidate drug and its target within the context of a whole, intact cell allows for more physiologically relevant data to be obtained. However, such assays are more complex than traditional biochemical assays as such facts as membrane permeability, cellular metabolism, cell variability, additional binding partners, and signal transduction must be considered. To help you navigate the complexities in designing cell-based assays, we have gathered insights collected over the years and compiled them to provide you with elements to consider when setting up your cell-based assays. After all, any assay, biochemical or cell-based, is only as good as its design.

Enhance your GTP measurements with this application note δ-opioid receptors (DOP) have become a major target for the development of new pain treatments. This application note will show you how to characterize pharmacological compounds easily through GTP binding assays: Measuring the level of Gi protein activation Using a CHO membrane model expressing delta opioid receptor Several case studies for the different classes of pharmacological compounds

Dive deeper into research on the GPCR signaling pathway β-arrestins are intracellular proteins that play an important role in GPCR signaling. Complexes formed between ligand-occupied GPCRs and β-arrestin lead to interaction with adaptor protein AP2. This interaction is followed by internalization of the receptors. HTRF technology is effective for studying the interaction between AP2 and β-arrestin2. Get your application note to discover: The applicability of the β-arr2 recruitment kit to a variety of compounds acting on β-arrestin2, and its ability to correctly rank pharmacological compounds (agonists and antagonists) How you can detect the β-arrestin2 / AP2 interaction for all classes of GPCRs Detailed experimental conditions and explanatory diagrams

Challenge the limits of binding kinetics studies This Note describes how binding kinetics studies can be enriched with a K on , K off approach by providing critical data on how the association and dissociation rates of a receptor-ligand couple can be assessed thanks to streamlined, no wash Tag-lite assays. Learn how to process and analyze the data, and discover how receptor binding kinetics offers significant insights into your compound’s mode of action. Features: Materials and methods for the experiment Data processing and result analysis Examples from our R&D

Discover a rapid and reliable tool for the identification and characterization of STING-targeting compounds. Quantifying human IFN-β using an HTRF-based approach This note demonstrates how using HTRF to quantify human IFN-β is a powerful and effective method for the characterization of STING agonists. It provides data demonstrating that the quantification of IFN-β using HTRF is well correlated with ELISA assays. This note also provides data about gene reporter assay correlation with our HTRF IFN-β assay.

Emerging pathways to neuroinflammation and neurodegeneration Neurodegenerative diseases, such as amyotrophic lateral sclerosis, Parkinson's disease, or Alzheimer's disease, occur as a result of neuroinflammation and neurodegenerative progression. For the moment, these diseases are incurable. However, as research progresses, many similarities between these diseases have been found at a sub-cellular level. Review the fundamentals of the neuroinflammation process Learn from a cutting-edge research report Detailed insight into neurodegenerative diseases

A Bruton Tyrosine Kinase (BTK) case study PROteolysis Targeting Chimeras (PROTACs) are one of the latest trending tools used in drug discovery. This note compiles experiments carried out to characterize PROTAC compounds using Bruton Tyrosine Kinase (BTK) as a model on Ramos cells. Both AlphaLISA and HTRF technologies were used in biochemical and cell-based contexts. Featured in this note: Assay principle and workflow Consistent results (data and graphs) Informative materials and methods, including references

Unlock the future of therapeutics with multispecific antibodies Discover the groundbreaking advancements in multispecific antibody research and their transformative potential in treating complex diseases. Download to: Understand the evolution from monospecific to multispecific formats and their clinical benefits. Explore various multispecific antibody structures, including Bispecific T-cell Engagers (BiTEs), ImmTACs, and DuoBodies. Learn advantages over monospecific antibodies: how multispecific antibodies offer improved specificity, efficacy, and reduced drug resistance. Delve into the FDA-approved multispecific antibodies and their applications in cancer, hemophilia, and eye diseases. Gain insights into the safety considerations and manufacturing complexities of multispecific antibodies. Discover ongoing research and future directions in multispecific antibody therapeutics. For research use only. Not for use in diagnostic procedures.

This application note shows you how to determine the relative affinity of different therapeutic antibodies to FcRn, and how it can be used to compare biosimilars. It contains experiments and data that investigate the specificity of the assays for binding the Fc portion of an antibody, and the binding performance of different subtypes. It features: Assay principle and workflow Consistent results (data and graphs) Informative materials and methods, including references

A comprehensive overview of the autoimmunity world This guide provides information on the diversity of the cell types and molecular pathways associated with autoimmunity, and the dysregulated immune responses associated with autoimmune diseases. The document focuses on Rheumatoid Arthritis, Systemic Lupus erythematous, Inflammatory Bowel Disease, Multiple Sclerosis, and Type I Diabetes. Features Pathogenesis underlying each of the autoimmune diseases listed above Cellular and molecular mechanisms generally associated with the diseases An overview of current and future treatments used for autoimmune diseases

A comprehensive overview of fibrosis development Fibrosis is a main contributor to a wide range of organ failures which stems from a mis-regulation of inflammation in response to injury. Fibrosis occurs when the long-term remodeling and removal of ECM suffers from a higher production than removal rate. This event is associated with chronic inflammation, which keeps myofibroblasts active for extended periods, thus making them and their ECM protein secretion the main effectors of pathological fibrosis. This guide is intended to help scientists and researchers appreciate and navigate the cellular and signaling actors of the development of fibrotic pathologies and disorders. Review the fundamentals of fibrosis as a process. A look at TGF-β1 signaling pathways as therapeutic approaches for fibrotic disorders. Insight into immune cell types involved and treating and reversing progressive.

A single White Paper for a review of immuno-oncology Immuno-oncology, the field which stimulates a individual's own immunity in order to fight cancer, has seen major advances since the 19th century and is now one of the most promising approaches against cancer. This White Paper provides you with relevant information related to: Immuno-oncology history Passive therapies - review Active therapies - review Immuno-oncology’s future

This technical note explores how pervanadate enhances tyrosine phosphorylation detection, enabling clearer insights into kinase activity and inhibitor efficacy in drug discovery research.

Over the last decade, therapeutic antibodies have emerged as the predominant class of new drugs. Assays to functionally characterize and evaluate antibodies can be used in the discovery process as well as during the development stages of antibody therapeutics. In this detailed case study, we show a suite of assays that can be used to compare an engineered antibody (adalimumab Fc Silent ™ ) to the original therapeutic antibody (Adalimumab). In this application note, discover how Alpha, HTRF and LabChip ® Protein Express assays can help when you are: Comparing the binding affinities of engineered antibody or original therapeutic antibody to the target or Fc receptors, or for readouts in functional cell-based assays Assessing binding capabilities after thermal stress, to check antibody stability Looking for critical data to build confidence in a biosimilar or an engineered therapeutic antibody

Improve your research on β-arrestin with HTRF™ technology The β-arrestins 1 and 2 play a central role in GPCR signaling pathways by regulating agonist-mediated GPCR signaling. The β-arrestins mediate both desensitization and resensitization processes, targeting many receptors for internalization by recruiting GPCRs to the AP-2 complex and clathrin internalization sinks. The involvement of β-arrestins appears to vary significantly depending on their expression, type, receptors, the ligands involved, and cell types. This application note presents a convincing demonstration of monitoring β -arrestin 1 and 2 expressed in various cellular models, using the HTRF total β-arrestin 1 and HTRF total β-arrestin 2 kits: Highly specific and independent detection of non-targeted β-arrestin expression level Ability to monitor the expression of endogenous and overexpressed β-arrestin 1 and 2 Potential applications for a variety of cell models

Investigate the dynamics of Interactions Kinetic values are of importance for characterizing protein-protein interactions. Studying them brings insight regarding macromolecule binding. Accessing kinetic values is therefore important. This application note dives deep into the topic to provide key guideline and experiments for determining true Kd values in protein-protein interaction assays with HTRF. Benefit from an expert's insight on: Handling HTRF data Determining Kd Investigating Prot-Prot Interactions

Studying mitophagy activation in relation to ubiquitin phosphorylation with HTRF Recent research studies have established that defective mitophagy constitutes one of the hallmarks of neurodegenerative disease onset, such as Alzheimer’s and Parkinson’s disease. Ubiquitin phosphorylation at serine 65 is proven to be a specific marker of ubiquitin-dependent PINK1/Parkin-mediated mitophagy induction, with PINK1/Parkin linked to Parkinson’s disease. In this application note we will show you how to investigate mitophagy activation in relation to phospho-ubiquitin (Ser 65) signaling using HTRF, a fast solution compared to standard assays and imaging. Features: Background on mitophagy, ubiquitin, and PINK1/Parkin Pathway involvement in neurodegenerative diseases Results showing the ability to test compounds with HTRF that may modulate the PINK1/Parkin pathway Materials and methods to perform these experiments

Discover the versatility and precision of Homogeneous Time-Resolved Fluorescence (HTRF) technology. Our HTRF portfolio offers a comprehensive range of assays designed to meet the needs of drug discovery and biomolecular research. This catalog includes: Phosphorylation cellular assays GPCRs assays Cytokine assays Immunity and cancer assays Biomarkers for main disease areas Tools for PPIs and assay development Kinase biochemical assays Epigenetics assays Biotherapeutics assays Custom Solutions HTRF™ technology in details For research use only. Not for use in diagnostic procedures.

Challenge kinase binding assays and win with HTRF™ technology The 3 application notes present a new HTRF kinase binding assay that combines the usage of a fluorescent inhibitor with a tagged kinase enzyme and its corresponding anti-tag-Europium labeled detection reagent. These 3 application notes will provide you with: Validation data for the 3 fluorescent tracers  Dasatinib-red, Staurosporine-red and Sunitinib-red Guidance for the determination of inhibitor constants: Kd and Ki Pharmacological characterization of known inhibitors and correlation with literature data Relevant case studies to help you revolutionize your assays