LANCE TR-FRET Assays

The measurement of protein phosphorylation is a useful tool for measuring the modulation of receptor activation by both antibodies and small molecules. CCR7 and CXCR2 receptors, which are expressed in immune cells and are therapeutic targets for disorders like lupus erythematosus, adult leukemia, lymphomas, chronic obstructive pulmonary disease (COPD), and sepsis. AlphaLISA ™ SureFire ® Ultra ™ and Alpha SureFire ® Ultra ™ Multiplex assays are automation-friendly, applicable to both small and large-scale screens, and can assess phosphorylation status in complex matrices. The LANCE Ultra cAMP assay is measures cyclic AMP (cAMP) produced upon modulation of adenylyl cyclase activity by G-protein coupled receptors (GPCRs). This application note demonstrates how the SureFire Ultra and LANCE Ultra cAMP assays can be used for measuring inhibitors to CCR7 and CXCR2 cell surface receptors using a cellular model system where these receptors are overexpressed in CHO cells. The assays were optimized to measure receptor blockage and assayed receptor activity modulation by detecting ERK and AKT phosphorylation status and cAMP modulation. For more details, download the application note!

Here, we demonstrate the utility and benefits of using LANCE Ultra TR-FRET assay technology for identifying and characterizing PD-1/PD-L1 binding inhibitors.

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

Epigenetic modifications in cancer The field of epigenetics has revealed revolutionary methods for understanding heredity. Epigenetic modifications can create new phenotypes that are inherited by later generations, without modifying the inherited DNA sequence itself. In this white paper, you’ll learn: The general description of the three types of epigenetic modifications How these three types of epigenetic modifications play diverse roles in tumorigenesis and cancer progression The promising future of epigenetics in cancer

Comprehensive Kinase and Antibody Selection for LANCE Ultra Assays Our comprehensive guide helps you identify and utilize kinases efficiently. It includes tested commercial kinase vendors and common aliases for easy reference. The guide lists the best performing ULight™ substrates for each enzyme, with signal to background (S/B) ratios provided under fixed conditions. Additionally, it helps you find the corresponding Europium-labeled anti-phospho antibody for your chosen substrate, essential for your LANCE Ultra assay. It offers two testing kits: the KinaSelect Ser/Thr Kit (TRF0300-C) with 5 ULight-labeled substrates and matching antibodies, and the KinaSelect TK Kit (TRF0301-D) with 1 ULight-labeled substrate and matching antibody. For research use only. Not for use in diagnostic procedures.

G protein coupled-receptors (GPCRs) are one of the most intensively studied drug targets, with up to one-third of all marketed therapeutics acting by binding to GPCRs. Therapies that target GPCRs include both agonist and antagonists that are used in the treatment of disease in nearly every major organ system and hold significance in several therapeutic areas, such as cardiovascular diseases, neurosciences, respiratory diseases, metabolic disorders, oncology, and rare diseases. We know the importance of furthering GPCR research - and we answer the call with application notes, white papers, and end-to-end solutions including reagents, instruments, and tools for all aspects of research into today’s most promising drug target. For research use only. Not for use in diagnostic procedures.

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

Guide to How Cellular Actors of the Tumor Microenvironment Challenge Immunotherapies Since the early 2000s, significant scientific breakthroughs have been made to better understand the complexity of the tumor microenvironment (TME) and its implication in shaping the response to cancer immunotherapy. It is now clear that the TME is one of the main hallmarks of cancer. Therefore, one of the major research challenges is identifying key pathways and factors in the TME that can be targeted to enhance the efficacy of immunotherapies. This guide focuses on the implication of cancer-associated fibroblasts (CAFs) and tumor associated macrophages (TAMs) on cancer immunotherapy. As master regulators of the tumor stroma, CAFs and TAMs define the immediate environment of cancer cells, and this parameter is critical for immunotherapeutic strategies that rely on immune cells having physical and chemical access to tumors. Download this guide to get insight on: The implication of CAFs and TAMs in immunotherapy How innovative tools can facilitate the study of tissue fibrosis and accelerate the identification of effective drugs

LANCE™ Ultra TR-FRET is a no-wash proximity assay technology that combines the benefits of time resolution (TR) with fluorescence resonance energy transfer (FRET) principles by using energy donor and energy acceptor fluorophore labels. For research use only. Not for use in diagnostic procedures.

The LANCE™ Ultra cAMP Assay Development Guidelines provide comprehensive instructions for the quantitative determination of cyclic adenosine monophosphate (cAMP) in cell lysate and cellular membrane samples. This guide details the assay principle, reagent preparation, protocols for cell preparation, and assay development workflow, emphasizing the assay’s sensitivity and robustness for high-throughput screening (HTS) applications. For research use only. Not for use in diagnostic procedures.

STAT3 is a particularly important therapeutic target since it is constitutively active in a range of cancer cells lines and human tumors. Therefore, fast and easy methods for screening of inhibitors of STAT3 activation are highly desirable.

This note presents a method to evaluate inhibitors of p38 kinase activity using LANCE. We are measuring energy transfer from a Eu-labeled antibody to a fluorophore-conjugated antibody binding to the GST portion of the fusion substrate

An in-depth review of molecular and cellular pathways The maintenance of proteostasis, the biological mechanisms that control the fate of a protein from synthesis to degradation, is ensured by complex and interconnected pathways controlling protein abundance, turnover, folding, functions, subcellular localization, and ultimate degradation. Impaired proteostasis can lead to the accumulation of damaged or aggregated proteins, or conversely, to excessive protein degradation. Proteostasis is thus involved in pathologies such as cancer, autoimmune diseases, and neurodegenerative disorders. This protein degradation booklet helps scientists and researchers appreciate and navigate the diversity of molecular pathways associated with protein degradation. Check out this exclusive guide that includes: Key information about the ubiquitin structure, function, and regulation The cellular and molecular understandings on proteasome and autophagy degradation machinery An overview of current therapeutic strategies, as well as the promising PROTAC approach to specifically target protein degradation

Your dream assay does not exist yet? Create it! This guide will provide you with tools to help you develop your very own protein:protein interaction assays for your research using HTRF™. Detailed steps, microplate plans, optimization procedures and data processing methods: let yourself be guided along the way to creating the assays for your experiments. Features Information relative to reagents, buffers and instrument handling Detailed guidelines and matrices to build, optimize and analyze your own assays Extensive example from our R&D to provide support