HTRF

Proteins and their interactions have a history of keen research and technological innovations. All scientists deciphering the protein interactions today are the heirs to this history and are now contributing to it, how well do you know it? Learn more in this infographic! For research use only. Not for use in diagnostic procedures.

Get the best out of your phosphorylation assays Combining phospho and total protein assays enables better analysis. This Application Note provides valuable guidelines for efficiently analyzing and interpreting results from such assay combinations. Check out all the tips and examples in features! Features Introduction to phospho and total protein assay relevance Tips for handling and interpreting data Examples from actual experiments

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.

What Are the Challenges and Solutions? All across the globe, AAVs are getting the attention of scientists and companies working in gene therapy, as they provide the right combination of characteristics that make them one of the most promising vector today. In 2021 only, the global gene therapy market was 4.1 billion USD, and AAV vector therapies made up more than 43% of that market value! Designing and manufacturing AAV vectors is complex, and to be successful, certain challenges must be addressed. This implies monitoring and optimtizing production for a thorough quality control process, including reliable ongoing characterization of process intermediates and the final product. Key features include: Overview and data on the gene therapy market Description of AAV vectors’ genomes and transfer Challenges associated with AAV vectors’ design and manufacturing Analytical methods for AAV vector quality control with a description of innovative no-wash AlphaLISA™ assays

In the dynamic landscape of pharmaceuticals, biotherapeutics research stands out as the fastest-growing sector. Scientists worldwide are on a quest for ever more efficient tools to develop therapeutic proteins. From lead selection to bioprocessing qualification, rigorous analytical characterization is essential. That’s where Revvity steps in. Our comprehensive line of no-wash tools revolutionizes biologics screening, mechanism-of-action studies, and biomanufacturing. With robustness and convenience at the forefront, we empower researchers to accelerate discoveries and drive innovation. Welcome to a new era of precision and efficiency in biotherapeutics research. For research use only. Not for use in diagnostic procedures.

The RAS family of genes, particularly K-Ras, plays a critical role in cancer biology. Despite its notorious difficulty as a therapeutic target, recent breakthroughs have brought new hope in treating cancers driven by K-Ras mutations. Our latest application note delves into innovative approaches to K-Ras inhibition, including small molecule inhibitors, synthetic lethality strategies, and PROTAC® molecules. We also showcase the high specificity and sensitivity of the no-wash HTRF™ K- Ras immunoassay, a cutting-edge tool that offers a reliable and precise method for evaluating K-Ras protein levels, outperforming traditional techniques. Discover how this assay can accelerate your research in targeting the elusive K-Ras.

Detecting and quantifying HCPs with automatable homogeneous immunoassays During biotherapeutic manufacturing and production, the host cells - a great majority of them being Chinese Hamster Ovary (CHO) cells – produce protein impurities that are called Host Cell Proteins (HCPs). Even if more than 99% of them are removed from the final product, the residual CHO HCPs can induce immunogenicity in individuals or reduce the potency, stability, or effectiveness of a drug. Therefore, to meet regulatory organizations’ guidelines (such as FDA or EMA) on CHO HCP levels, biopharmaceutical companies spend significant amounts of money on tools and strategies for their detection. Illustrated with robust results, this Application Note explains the many ways in which HTRF™ and AlphaLISA™ kits can improve the workflow for CHO HCP detection: Wide antibody coverage Compatibility with most commonly used buffers No cross-reactivity between CHO HCP detection and drug substance Good dilutional linearity and antigen spike recovery

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

With over 200 different types of cancer, management relies on a variety of techniques such as chemotherapy, radiotherapy, and surgery. These types of therapies can be associated with severe side effects, and finding safer and more effective treatments is a priority in cancer therapeutics research. One approach that has shown huge potential is monoclonal antibody-based cancer therapeutics. In this white paper we explore this exciting area of anti-cancer research, covering mechanism of action, development, and challenges in monoclonal antibody-based therapeutics, including antibody-drug conjugates and multispecific antibodies.

Compound MoA and potential cytotoxic effects deciphered thanks to immunoassays and cell viability assay The attrition of drug molecules occurs at various stages in the development process, and most early-phase failures are attributed to safety and toxicity issues. Considering the economic impact of early project termination, the biggest challenge comes from selecting the most potent and selective drug compounds while assessing their potential toxic side effects. In this application note, you’ll discover how to easily combine AlphaLISA™, HTRF™, or AlphaLISA™ SureFire® Ultra™ immunoassays with the ATPlite™ 1step cell viability assay to simultaneously, and in a single sample, study the efficacy of drug compounds on disease pathways while identifying possible cytotoxic effects. Recommendations Detailed workflows Case studies on various cell lines and different markers using HTRF, AlphaLISA, or AlphaLISA SureFire Ultra immunoassays together with the ATPlite 1step cell viability assay

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 NAFLD knowledge with this booklet. NASH disease is complex and follows many development pathways. This booklet provides you with critical information regarding NAFLD and more specifically about NASH progression. Review the fundamentals of NAFLD and NASH learn from an important research report Benefit from additional content to help your NASH research

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

CDK9 is an attractive target for cancer therapeutics due to its crucial role in transcription regulation, particularly of short-lived anti-apoptotic proteins such as MCL-1 and XIAP which are critical to the survival of cancer cells. One approach for specifically degrading CDK9 is using a proteolysis targeting chimera (PROTAC). In this application note, you will discover: The rapid and strong effect of CDK9 PROTAC molecule Thal-SNS-032, demonstrating specificity in CDK9 degradation How HTRF ® and AlphaLISA ® SureFire ® Ultra ™ are used to measure protein levels, with each tool showcasing the ability of PROTACs to rapidly degrade a targeted protein making it a unique tool in the fight against cancer

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

With the increased understanding of molecular and cellular medicine, more specific and efficient gene transfer vectors are now producing clinical results. 20 years after the first clinical trials, gene therapy is considered one of the greatest scientific success stories of the 21st century. While the Covid-19 crisis has disrupted the evolution in this promising field, gene therapy is expected to recover and continue its growth in the future. Download this complete industry report to learn more about the gene therapy market trends and latest advances. It includes: Gene therapies in the global pipeline and major actors in the field A description of gene therapy development and vectors used The challenges concerning gene therapy manufacturing The promising future of gene therapy

The definitive guide to setting up a successful cytokine assay Many therapeutic areas require an understanding of cytokine release. When preparing for a cytokine assay, many underappreciated parameters (e.g. sample handling, cell culture format, sample dilutions) can in fact greatly impact the performance of the cytokine detection. This guide reviews the latest knowledge surrounding the proper use of HTRF cytokine assays. A review of the key terms and definitions in cytokine detection A list of optimization steps Recommendations for data analysis

Ask real blood for real responses Fresh blood is the model of choice to study drug immunotoxicity and predict adverse effects. Written in collaboration with Blood Assay Solutions, this note provides guidelines for fresh blood cytokine quantification. Discover the power of our cytokine portfolio for your research. Features Step by step protocols for fresh blood cytokine quantification in your research Examples of pathway stimulation assays (TCR, TLR …) Comparison beteen fresh blood and PBMCs

Evaluation of the therapeutic profile of anti-oncogene compounds in various cell lines with AlphaLISA™ and HTRF™ KRAS is a proto-oncogene known to be mutated in many cancer subtypes, inducing uncontrolled proliferation and cell metabolism changes. Like most small GTPases, KRAS will bind to GDP in its inactive form or to GTP in its active form. KRAS G12C is one of the most commonly found mutant forms in cancers, and leads to a permanently active state of KRAS. The upregulation of KRAS interaction with the exchange factor SOS1 leads to cancer phenotypes. Reducing KRAS activity and associated pathways could control the biological processes involved in cancer growth. Furthermore, it is well known that KRAS induces activation of mitogen-activated protein kinase (MAPK), thus playing a central role in human cancers. This application note provides a convincing demonstration of the reliability of the AlphaLISA and HTRF KRAS portfolios to evaluate compound in vitro therapeutic profiles in a cellular context: Determine the effects of KRAS and SOS1 inhibitors in different human cancer cell lines Discriminate the cellular action of KRAS-targeting compounds and evaluate their effectiveness in modulating KRAS downstream pathways.

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