June 27, 2018


The Agilent pH-Xtra™ Glycolysis Assay allows direct, real-time, kinetic analysis of extracellular acidification rates (ECA/ECAR). BioTek offers a dedicated filter cube assembly, pre-programmed Gen5™ protocol, and user tutorial specifically optimized for use with the lifetime signal acquisition of the assay on the BioTek Cytation 1, Cytation 5, or Synergy H1, or Synergy Neo2  instruments equipped with TRF capability.

8040588 cube
Filter Cubes:
P/N 8040588 or P/N 8040594 (Cytations, Synergy H1)
P/N 1035124 CUBE 124 PH-XTRA GLYCOLYSIS ASBY (Synergy Neo2)
Gen5 Data Analysis Protocol: AgilentpHX.prt
Note: The User Tutorial and Data Analysis Protocol are compatible with Gen5 v3.04 and higher software. Existing Gen5 users can upgrade their software for free at: https://btresource.force.com/CRC/s/

Example data of a glucose oxidase

Figure 1. Example data of a glucose oxidase titration performed on the Synergy H1 illustrates the detection principle of the Agilent pH probe: higher lifetime signals (left) equate to lower sample pH values (middle). pH values can then be converted to H+ and ECAR using Gen5 software (right).

A summary of the AgilentpHX.prt procedure, plate

Figure 2. A summary of the AgilentpHX.prt procedure, plate layout, and data reduction steps as shown from within Gen5 v3.04

Top level implementation steps of the Agilent pH-Xtra Glycolysis Assay toolkit for

Figure 3. Top level implementation steps for the Agilent pH-Xtra Glycolysis Assay with the BioTek Cytation 1, Cytation 5, Synergy H1, or Synergy Neo2 instruments. The toolkit is designed for models equipped with Time Resolved Fluorescence (TRF) capability.