Agilent Seahorse XF Glycolytic Rate Assay Single-File and Multi-File Report Generator FAQs

Document created by courtneynadeau Employee on Oct 15, 2019Last modified by courtneynadeau Employee on Oct 16, 2019
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What rate measurements are used to calculate the parameters in the Report Generator?

Parameter equations are described in Table 1 on page 6 and Table 2 on page 17 of the Single File RG user guide.

Parameter equations are described in Table 1 on page 6 and Table 2 on page 21 of the Multi-File RG User Guide.


Incompatible elements with the XF Glycolytic Rate Assay The following items are not supported with the XF Glycolytic Rate Assay:

  • Sensor cartridges for the XF24 and XF24-3 Analyzer.
  • Spheroid, Islet, and V28 plate types.


How do I remove outlier wells in the Report Generator?

Outlier assay wells must be turned OFF or reassigned in Wave 2.4 before Excel export. See “Exclude Outliers/Groups from Analysis” on page 23 or the Wave user guide for more information.


I need to change Buffer Factor for a group, where can I find this information?

Buffer Factor values are automatically imported from each Excel File per group. Modifying the Buffer Factor in the Multi-File XF Report Generator is not supported presently. To change the Buffer Factor value for a group, open the assay result file in Wave 2.4 software, change the Buffer Factor value of the assay media and the background wells, and re-export the data file to Excel.


Can I use the Excel file exported from the XFp, XF96 Analyzer, or an earlier version of Wave software?

This Multi-File XF Report Generator supports import and analysis of Excel files exported from Wave 2.4 (Desktop or Controller) or later. Excel files exported from the XFp Analyzer, XF96 Analyzer, XF Reader software, or earlier versions of Wave software are not supported.


If you receive an error message about Instrument Protocol (XFe96; XFe24; XF96 only)

Custom Cycles are not part of the standardized assay template for the XF Glycolytic Rate Assay and not supported in Report Generator analysis. A Custom Cycle refers to an axtra 'Mix' or 'Wait' command step in the Instrument Protocol of an assay. Please contact Agilent Seahorse Technical Support if you have any additional questions regarding Custom Cycles.


Can I use baseline rate data (%) to calculate assay parameters?

Normalized (or nonnormalized) ECAR data is used for parameter calculations, parameter calculations using baseline rate data (%) is not supported currently.


Will I see Warning message(s) when the % PER from Glycolysis is less than 50% and less than 10%

When the % of PER from Glycolysis for a selected group is less than 50% or less than 10%, the Report Generator will display a warning message in the legend on the Summary Printout tab (below the Normalize button). For more information, see “Advanced Options” on page 19.


More FAQs: Cell Analysis FAQs and Troubleshooting


Keywords: Glycolytic Rate Assay, phenol-red free, Buffer Factor, HEPES, Report Generator, Warning Message, % PER, Report Generator Error, Outlier, Parameter Equations, Seahorse, XFe96, XFe24, XFp, FAQ