Agilent Technologies Seahorse XFp, User Manual

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16 Agilent Seahorse XFp Glycolytic Rate Assay Kit User Guide
Assay Workflow
Prepare stock compounds
Use compounds the same day they are reconstituted. Do not
refreeze. Discard any remaining compound.
1
Remove one foil pack from the kit box, then open the pouch
and remove the Rot/AA (red cap) vial and one 2-DG (green
cap) vial.
2
Tap down the vials to ensure powder is on the bottom of the
tube before opening the vials.
3 Resuspend each component with prepared assay medium as
described in
Table 2 with a p1000 pipette. Tap down the
Rot/AA to ensure the powder is on the bottom of the tube.
Vortex
~
1 minute to ensure that compounds go into solution.
Dilute compounds
Table 3 describes how to prepare compounds dilutions to load
the cartridge. Please note that if different starting assay
volumes or port volumes are used, adjust compounds
concentrations in order to obtain the recommended final
concentrations in the well.
Table 2 Stock solutions

Compound
Volume of assay
medium Resulting stock
concentration
Rot/AA 216 µL 25 µM
2-DG 300 µL 500 mM
Table 3 Compound preparation for running the XFp Glycolytic Rate
Assay on an XFp Analyzer
Agilent
Seahorse XFp
Port A

Rot/AA
(Final well)
(µM) Stock volume
(µL) Medium
volume (µL) 10X (Port)
(µM)
0.5 60 240 5
Port B

2-DG
(Final well)
(mM) Stock volume
(µL) Medium
volume (µL) 10X (Port)
(mM)
50 300 0 500