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Jolanta Vidugiriene, Ph.D. - "Cellular Metabolism Assays: Revealing the link between metabolic changes and cell function"

Senior Scientist at Promega Corporation
When Oct 23, 2017
from 02:30 PM to 03:30 PM
Where Tigem Auditorium "Vesuvius"
Contact Name
Contact Phone 081-19230659
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Abstract
The growing trend towards understanding the role of cell metabolism in cancer, immunology, obesity, diabetes, and neurodegenerative disease has presented specific challenges in developing rapid and reliable methods for measuring the changes in metabolic pathways. Current technologies are dependent on analytical methods or enzyme-coupled absorbance/fluorescent assays and often require laborious sample preparation and/or lack sensitivity and dynamic range for rapid and convenient metabolite detection directly in 96-well plates.
Here we introduce luminescence metabolite detection assays (e.g. glucose uptake, glucose, lactate, glutamate, NAD(P)/NAD(P)H detection) and discuss how those assays provide valuable information about metabolic reprograming of cancer cells, allow monitoring activation of T cells or evaluation of insulin sensitivity in primary adipocytes.
Changes in metabolic reprograming are often associated with increased ROS levels, and deregulated redox homeostasis is a common feature of cancer cells. We will briefly review the performance of luminescent GSH/GSSG and H2O2 assays and discuss how multiplexing of different assay chemistries and incorporation of normalization readouts, for example viability measurements, into experimental workflow improves data quality and allows you to obtain more information from the same set of samples.  In addition, since metabolic perturbation frequently coincides with alterations in autophagic activity, a reporter-based luminescent assay for microplate determination of autophagic flux will be discussed.

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