Cell Based Assays

The Greatness of Glycogen: A Central Storage Molecule in Energy Metabolism

Posted on by Simon Moe

Introduction

Glycogen is a fundamental molecule in energy metabolism, serving as the critical storage form of glucose that supports cellular health and energy homeostasis. As a polysaccharide, glycogen is essential for maintaining stable energy levels, particularly during periods of fasting and physical exertion. This article will examine glycogen’s synthesis, storage, and utilization, along with its broader significance in human health and disease. Understanding glycogen’s role can provide valuable insight into energy regulation and metabolic health.

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The Benefits of BCAAs: Branched-Chain Amino Acids in Health and Disease

Posted on by Simon Moe

Introduction

Branched-chain amino acids (BCAAs) are essential nutrients that play a significant role in muscle metabolism and overall health. Comprised of leucine, isoleucine, and valine, BCAAs cannot be synthesized by the body and must be obtained through diet. Recent research has highlighted how the metabolic pathways are influenced by BCAAs, such as their ability to activate mTOR signaling, which is vital for muscle protein synthesis (Choi, 2024). Beyond muscle growth, BCAAs may support cognitive function and metabolic health, with ongoing research exploring their broader benefits in disease management. This article explores the diverse roles of BCAAs and their impact on health and diseases

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The Brilliance of BHB: A Key Ketone Body in Metabolic Health

Posted on by Simon Moe

Introduction

β-Hydroxybutyrate (BHB), the most abundant ketone body, is a crucial molecule that sustains energy production during periods of glucose deprivation. Whether you are fasting, adhering to a ketogenic diet, or simply interested in metabolic flexibility, BHB offers key insights into how our bodies adapt to alternative energy sources. This article will delve into how BHB is produced, the diverse roles it plays, and its implications for health and disease.

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Visualize Protein:Protein Interactions with Bioluminescence Imaging

Posted on by Simon Moe

If you’re familiar with bioluminescence, you’ve probably used it in plate-based experiments to track various biological processes. You understand it provides distinct advantages over traditional fluorescence assays, particularly when it comes to sensitivity. However, there’s always that one nagging question: how representative is the signal on a cell-to-cell level?

Traditional approaches to decipher cell-to-cell signal rely on complex, time-intensive measures that only approximated the findings acquired through bioluminescence. That’s where the GloMax® Galaxy Bioluminescence Imager comes in. This new tool will enhance your ability to visualize proteins using NanoLuc® technology, going beyond simple numeric outputs to reveal what’s happening in individual cells.

NanoLuc® technology is well-known for its ability to assist in detecting subtle protein interactions in complex biological systems. This bright luminescent enzyme enables a much broader linear range than fluorescence, improving detection of small changes in protein activity, such as proteins interacting. Microplate readers measuring NanoLuc® assays rely on signal generated from many cells. This results in an approximation of what is occurring biologically. Truly validating those luminescent readings within a cell population has been challenging—until now. The GloMax® Galaxy allows you to perform bioluminescence imaging, moving beyond the numbers, offering the power to visualize protein interactions directly.

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Immunometabolism: The Dynamic Interplay of Cytokines and Metabolites

Posted on by Kendra Hanslik

Immunometabolism is the study of how metabolic processes influence immune cell functions and how immune responses, in turn, shape cellular metabolism. This field examines the roles of cytokines and metabolites, which act as signaling molecules and energy sources, respectively. Cytokines can trigger or modulate metabolic pathways in immune cells, affecting their activation, growth, and response capabilities. Similarly, metabolites provide the necessary energy and building blocks that enable immune cells to proliferate, function optimally, and sustain their activity during immune responses. This dynamic interplay is crucial for maintaining health and combating disease. Together, cytokines and metabolites orchestrate a complex network that links metabolic health with immune competence on a systemic and cellular level. This blog discusses how cytokines and metabolites not only influence but also drive immune cell functions, revealing new avenues for therapeutic interventions across a range of diseases.

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Rethinking Cell Proliferation Assays

Posted on by Simon Moe

In the field of cancer research, accurately measuring cell proliferation is crucial for assessing the efficacy of therapeutic agents. This is particularly difficult with CDK 4/6 inhibitors, which arrest cells in the G1 phase without stopping their growth. This continued growth can skew results from proliferation assays which rely on factors that naturally scale with cell growth. These include mitochondrial activity (ATP levels), total cell protein, or mRNA as measured through the PRISM molecular barcoding strategy. Even though these cells are not dividing, the increase in these measurements can misleadingly suggest active proliferation.

There is growing awareness among researchers of these challenges.  A recent study highlights these limitations by demonstrating the discrepancies that arise when using metabolic assays to assess cell proliferation after treatment with drugs that induce cell cycle arrest. This blog post delves into the study’s implications and demonstrates how one of Promega’s latest developments is poised to address these challenges.

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Decoding the NAD+/NADH Ratio and Its Crucial Role in Cell Health

Posted on by Kendra Hanslik

Nicotinamide adenine dinucleotide (NAD) exists in two forms in the cell: NAD+ (oxidized) and NADH (reduced). This molecule plays a pivotal role in metabolic processes, serving as a key electron carrier in the redox reactions that drive cellular metabolism. The balance between these two forms, commonly expressed as the NAD+/NADH ratio, is crucial for maintaining cellular function and the intracellular redox state. This article explores the significance of this ratio, how it impacts cellular processes, and the consequences of NAD+/NADH ratio dysregulation.

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Connecting Synaptic Gene Polymorphisms to Parkinson’s Disease

Posted on by Kendra Hanslik
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Neurodegenerative disorders represent a significant and growing concern in the realm of public health, particularly as global populations age. Among these, Parkinson’s disease (PD) stands out due to its increasing prevalence and profound impact on individuals. Characterized by the progressive degeneration of motor functions, PD is not just a health challenge but also poses substantial socio-economic burdens. While the etiology of Parkinson’s disease is far from simple, current research efforts elucidating its causes, mechanisms, and potential treatments illustrate the critical nature of this neurodegenerative disorder in today’s healthcare landscape.

In the clinic, Parkinson’s disease is often diagnosed as either sporadic or familial. Familial PD has a clear genetic basis, typically passed down through families, while sporadic PD, comprising about 90% of cases, occurs in individuals without a known family history of the disease. The exact cause of sporadic PD is not fully understood but is believed to be due to a combination of genetic predispositions and environmental factors. In contrast, the factors involved in familial PD are more thoroughly understood, offering insights into the molecular mechanisms underlying PD pathogenesis.

Polymorphisms and Parkinson’s Disease Susceptibility

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Cell-Based Target Engagement and Functional Assays for NLRP3 Inhibitor Profiling Help Identify Successes and Failures

Posted on by Kari Kenefick

Identifying Inflammasome Inhibitors: What’s Missing
The NLRP3 inflammasome is implicated in a wide range of diseases. The ability to inhibit this protein complex could provide more precise, targeted relief to inflammatory disease sufferers than current broad-spectrum anti-inflammatory compounds, potentially without side effects.

Studies of NLRP3 inflammasome inhibitors have relied on cell-free assays using purified NLRP3. But cell-free assays cannot assess physical engagement of the inhibitor and target in the cellular micro-environment. Cell-free assays cannot show if an NLRP3 inhibitor enters the cell, binds the target and how long the inhibitor binding lasts.

Cell-based assays that interrogate the physical interaction of the NLRP3 target and inhibitor inside cells are needed.

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Scaling Up to Measure 40,000 Data Points a Day with GloMax® Microplate Readers

Posted on by Jordan Nutting

Traditional approaches for protein degrader compound screening like Western blotting can be laborious, time consuming and cannot be streamlined with automation. By implementing a high-throughput, automated workflow that uses our CRISPER/Cas9 knock-in cell lines, live-cell bioluminescent assays and sensitive GloMax® Discover microplate readers, our custom assay services offer protein degradation profiling at an accelerated rate.  

To do this, we collaborated with HighRes® Biosolutions, to develop an automated system that can screen up to 100 384-well plates each day, generating roughly 40,000 data points with minimal hands-on work.

Learn how bioluminescent tools like HiBiT and NanoBRET™ technology can help you answer key questions in your targeted protein degradation research.

An important step of building this system is to integrate four GloMax® Discover microplate readers into the automated system using instrument’s built-in SiLA2 communication driver. The driver software makes it easy to connect the microplate readers with HighRes® Biosolution’s robotic components.

Check out our setup in the video below.

See how we’ve integrated GloMax® Discover microplate readers into a high-throughput automated system for profiling protein degraders in live cells.
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