Seahorse XF24, Руководство по обучению

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Basic Procedure
Cell Density with the XF Glycolysis Stress Test
The XF Glycolysis Stress Test is run with four different cell densities to determine the optimal cell density
to use in your XF assays. In a typical cell density optimization assay, only three basal measurements
need to be taken to determine the optimal cell seeding density. However, for the purposes of providing
richer data for discussion, we will run the XF Glycolysis Stress Test and inject (A) Glucose, (B) Oligomycin
and (C) 2-Deoxy-D-glucose (2-DG). In this experiment, we will use 1.0 µM oligomycin, although the
optimal concentration may be different (as is investigated in the next experiment).
Plate Layout:
Injections:
Port A: Glucose - 10 mM final concentration in the well
(80 mM stock)
Port B: Oligomycin - 1.0
µM
final concentration in the well
(9
µM
stock)
Port C: 2-DG – 100 mM final concentration in the well
(1 M stock)
Cell #
10 K
20 K
40 K
80 K
22