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Agilent Technologies, Inc.

Seahorse XF Glycolytic Rate Assay Kit

The XF Glycolytic Rate Assay provides a precise measure of glycolysis in live cells, revealing transient responses and rapid metabolic switches not discernible in endpoint lactate assays. Run on a Seahorse XF Analyzer, this assay quantifies proton efflux specific to glycolysis. This assay should be performed using XF pH-defined media.

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  • Calculates and subtracts mitochondrial-produced acidification for a precise measure of glycolytic rate
  • Detects metabolic switching (such as Warburg effect) by reporting relative activity of mitochondrial respiration and glycolysis pathways
  • Measures before and after effects of metabolic modulators on glycolytic rates within a single assay well
  • This assay is best performed with Seahorse XF DMEM medium, pH 7.4 (Cat No. 103575-100) or Seahorse XF RPMI medium, pH 7.4 (Cat No. 103576-100). For other assay media options, refer to XF buffer factor protocol quick reference guide.
  • Kit contains Rotenone/Antimycin A mix to block mitochondrial activity (for calculation of mitochondrial acidification) and 2-DG (to inhibit glycolysis) as an internal control
  • Single-use vials assure quality and quantity every time and are now easier to open
  • Glycolytic rate, percent of proton efflux due to glycolysis, and other key metrics are calculated by the Report Generator
  • Standard protocol uses phenol red free Base Medium and 5mM Hepes for higher dynamic range and data quality
  • For use with an XFe96, XF96, or XFe24 Analyzer. Not compatible with XF24 and XF24-3 analyzers
  • For the kit compatible with Seahorse XFp Analyzers, please see the Seahorse XFp Glycolytic Rate Assay Kit

How it works

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Provides real-time measurements of glycolysis

This assay detects glycolysis modulation in minutes in live cells. Researchers use this important assay to advance their understanding of immune cell activation, cancer cell metabolism and cell proliferation.  The assay reports multiple key parameters, such as basal glycolysis (a physiological rate) and compensatory glycolysis.  Compensatory glycolysis is what cells can achieve when mitochondrial respiration is inhibited.  This parameter is indicative of cellular ability to manage energy demand under stress.

Learn More



Download the Agilent Seahorse XF Assay Kits & Reagents Overview
Download the new Media Selection Guide
Download the Seahorse Plastics Brochure
Download the Seahorse XF Instrument Selection Guide


Agilent Technologies, Inc.

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Measure mitochondrial respiration and glycolysis as well as ATP production rate of live cells.

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