How to Integrate Cellular Metabolism Assays Into Your Research: Considerations and Challenges
- Critical factors in experimental design: sensitivity, linear range, cell media, and controls
- Strategies for simpler sample preparation of cells, media and tissues
- Getting more information from individual samples by quantitating multiple metabolites and multiplexing with a viability and normalization assay
- Monitoring changes in metabolite levels over time during T cell activation and growth of cancer cell lines
Summary
Cellular metabolism comprises a dynamic network of pathways that undergo reprogramming in response to external and internal signals. This reprogramming has vital roles in proper cell function and also in many disease states, making cancer metabolism and immuno metabolism important areas for research and drug discovery. Metabolite levels provide valuable information about pathway activity, and plate-based coupled-enzyme reaction assays can provide a rapid method for measuring specific metabolites. However, current plate-based metabolite detection assays can pose challenges especially with sample preparation, reproducibility and throughput.
In this webinar we will discuss new plate-based bioluminescent assays for measuring the key metabolites glucose, lactate, glutamine and glutamate, and demonstrate using these assays to study glycolysis and glutaminolysis, two major fuel-metabolizing pathways. We will discuss factors to consider when designing an experiment, including assay sensitivity and linear range, as well as choice of sample preparation method.
Speaker
Donna Leippe, PhD
Senior Research Scientist
Dr. Donna Leippe has spent over 18 years developing technologies and leading research projects as a Senior Research Scientist in R&D at Promega. She is a member of the Assay Design group and focuses on developing bioluminescent technologies for cellular metabolism research. This has included assays for measuring cellular cofactors (NAD/NADH-Glo™ and NADP/NADPH-Glo™ Assays) and multiple cellular metabolites (Lactate-Glo™, Glucose-Glo™, Glutamate-Glo™ and Glutamine/Glutamate-Glo™ Assays). Donna joined Promega after completing her graduate and postdoctoral work at the University of Wisconsin-Madison, where she studied Viral Immunology at the Institute for Molecular Virology and the McArdle Laboratory for Cancer Research.