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Agilent Seahorse XFp Glycolytic Rate Assay Kit User Guide
Introduction
Figure 2
Principle of the Agilent Seahorse XFp Glycolytic Rate Assay.
Energy is produced by two pathways in the cell - glycolysis,
and mitochondrial respiration. In the breakdown of glucose to
lactate during glycolysis, protons are extruded into the
extracellular media which is detected by the XFp Analyzer as
ECAR. Additionally, mitochondrial TCA activity produces CO
2
,
which hydrates and acidifies the media. By inhibiting
respiration (OCR) during the assay with the complex I and
complex III mitochondrial inhibitors (Rot/AA), the rate of
proton efflux from respiration can be calculated and removed
from the total proton efflux rate, giving the glycolytic proton
efflux rate (glycoPER). To confirm pathway specificity, 2-DG, an
inhbitor of glycolysis is injected to stop glycolytic acidfication.
Summary of Contents for Seahorse XFp
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