Neuroscience Research Reagents

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.

Discover Alpha SureFire ®   Ultra ™ assays, the no-wash cellular kinase assays leveraging Revvity's exclusive bead-based technology and sandwich immunoassays for detecting phosphorylated proteins in cells. Offering a quantitative alternative to Western Blotting, Alpha SureFire assays are automation-friendly, easily miniaturized, and proficient in detecting both endogenous and recombinant proteins. Explore our comprehensive portfolio of SureFire Assays, designed to help you elevate and expedite your drug discovery journey.

Quantifying H3K27ac Levels with AlphaLISA Assay In this technical note, you will discover how AlphaLISA immunodetection assay easily detects alterations in acetylated histone H3 lysine 27 (H3K27ac) levels within cellular extracts.

Decoding Histone Modifications: A Targeted Assay for H3K4me2 In this study, you will find details about an optimized assay designed to quantify changes in H3K4me2 levels following treatment with sodium butyrate and Trichostatin A (TSA), both of which are non-selective histone deacetylase (HDAC) inhibitors. Using a standardized histone extraction procedure, we detect the target mark by employing a biotinylated anti-Histone H3 (C-terminus) antibody and AlphaLISA Acceptor beads conjugated to an antibody specific to the mark.

Measuring Lysine 27 Tri-Methylation Demethylation in Histone H3 with AlphaLISA In this technical note, you will discover how this AlphaLISA immunodetection assay detects the demethylation of a biotinylated Histone H3 (21-44) peptide that is tri-methylated at lysine 27.

AlphaLISA Assay for Mono-Methylation of Histone H3 at Lysine 4 In this technical note, you will discover how this AlphaLISA immunoassay detects the mono-methylation of a biotinylated Histone H3 (1-21) peptide at lysine 4.

This guide outlines further possible optimization of cellular and immunoassay parameters to ensure the best possible results are obtained.

This application note explores how the AlphaLISA™ SureFire® Ultra™ technology reveals the intricacies of TREM2/DAP12 signaling in neuroinflammation.

The definitive guide for setting up a successful AlphaLISA SureFire Ultra assay Several biological processes are regulated by protein phosphorylation. It is, therefore, no surprise that the dysregulation of protein phosphorylation is implicated in a relatively large number of diseases. AlphaLISA SureFire Ultra assays provide a robust and reliable method for quantifying a targeted phosphorylation event in cell-based experiments. This guide contains tools and data helpful for you to perform your assays using AlphaLISA SureFire Ultra: A detailed description of the assay and its options A thorough investigation of assay conditions to obtain the optimal response from the chosen modulator and cell line A list of optimization steps to provide a sufficient assay window and produce the strongest results possible

Homogeneous Assay for Tri-Methylated Histone H3 Lysine 27 in Cellular Extracts In this technical note discover how AlphaLISA ™ assay tracks variations in cellular extract levels of tri-methylated histone H3 lysine 27 (H3K27me3).

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

Immunoassays are a mainstay for the quantification of a variety of biomolecular analytes in drug discovery, drug development, and life sciences research. AlphaLISA® proves advantageous to ELISAs, offering a novel, homogenous immunoassay technology that eliminates wash steps. Using the JANUS® Automated Workstation in combination with AlphaLISA provides a solution to easily preparing assays that can be tailored to the needs of the laboratory. Moreover, the JANUS workstation can be easily set-up to process different assay types in a multi-user, multi-assay environment, thus providing flexible automated workflows.

AlphaLISA™ technology is a highly sensitive, easy-to-use, and reproducible method for detecting and quantifying molecules in various biological matrices. It works by using streptavidin-coated Donor beads and biotinylated anti-analyte antibodies. When these come into close proximity, the excitation of the Donor beads at 680 nM triggers an energy transfer cascade in the Acceptor beads, generating a sharp emission peak at 615 nM. However, some cell culture media contain high levels of biotin, which can interfere with AlphaLISA and other assay technologies that rely on a streptavidin-biotin binding event for detection. High levels of free biotin in the sample matrix can result in a decrease in total counts, lower signal to background ratios, and reduced AlphaLISA assay detection limits. To mitigate this, AlphaLISA biotin-free kits have been developed. This application note demonstrates the value of using AlphaLISA biotin-free kits to reduce the effects of biotin interference in sample and standard preparations.

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

Protocol for brain sample analysis with HTRF In this Technical Note, you will find tips we have complied to help ensure you are able to make the most of your brain sample. The Insider Tips you will find includes: Lysis step Determination of protein concentration Running of the optimized HTRF assay

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!

AlphaLISA and LANCE (TR-FRET) biomarker assays can be used to measure ECM-associated protein modulation caused by human transforming growth factor-beta (TGF-β) induction of EMT in a 3D Spheroid model of human prostate carcinoma.

Scientists around the world have been working at warp speed to find the best ways to treat and prevent SARS-CoV-2 infection. In contrast to rapid development of effective vaccines, treatments for COVID-19 remain somewhat elusive. Multiple approaches to identify potential therapeutics targeting SARS-CoV-2 infections are currently being pursued. Most of the treatments fall into one of three categories: antivirals, antibodies, or protein-protein interaction inhibitors. In the following literature review, we have highlighted the various approaches that researchers have adopted to identify potential new therapeutics and the challenges they face when it comes to the latest variants.

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

Cytokines are implicated in the pathology and outcome of disease states across all therapeutic areas. When preparing to run cytokine assays, it is key to consider a multitude of factors that impact the assay performance, such as sample dilutions and cell culture format. Utilize this guide to optimize your AlphaLISA ™ cytokine assays with in-depth data and information!

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

Due to limitations driven by circulatory half-life and drug target bioavailability, injected biologics often require the injection of high doses, which can result in patient discomfort, unwanted side effects, a limited therapeutic window, and higher costs. To sidestep these pain points, Tenza has engineered probiotic microbes to synthesize and deliver protein therapeutics directly to the target tissue. The functional activity of the secreted protein biologic is assessed by its binding to a target protein relevant to its therapeutic indication. The pre-existing assay format for testing functional activity was a standard ELISA, which had limited dynamic range and throughput, required large sample volumes, and involved multiple tedious wash steps. Download this application note and discover how the authors switched to a custom developed AlphaLISA ® assay to overcome the limitations they had observed in the use of their ELISA assays.