Related Manuals for Agilent Technologies Seahorse XFp
Summary of Contents for Agilent Technologies Seahorse XFp
- Page 1 Agilent Seahorse XFp Glycolytic Rate Assay Kit User Guide Kit 103346-100 Agilent Technologies...
- Page 2 Further, to the max- agreement and written consent from imum extent permitted by applicable Agilent Technologies, Inc. as governed by C A U T I O N law, Agilent disclaims all warranties, United States and international copyright either express or implied, with regard laws.
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Page 3: Table Of Contents
Kit Shipping and Storage Additional Required Items Assay Workflow Day Prior to Assay Day of Assay Running the Assay Analysis Using the Agilent Seahorse Glycolytic Rate Assay Report Generator Frequently Asked Questions Agilent Seahorse XFp Glycolytic Rate Assay Kit User Guide... - Page 4 Agilent Seahorse XFp Glycolytic Rate Assay Kit User Guide...
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Page 5: Assay Background
Assay Background For use with an Agilent Seahorse XFp Extracellular Flux Analyzer. The Agilent Seahorse XFp Glycolytic Rate Assay is an accurate and reliable method for measuring glycolysis in cells. Together with a Seahorse XFp Analyzer, the Seahorse XFp Glycolytic... - Page 6 The conversion of glucose to lactate results in a net production and extrusion of protons into the extracellular medium. (See Figure 2 on page 8.) Cells may also use glucose and the other fuels present within Agilent Seahorse XFp Glycolytic Rate Assay Kit User Guide...
- Page 7 The resulting decrease in PER provides qualitative confirmation that the PER produced prior to the injection is primarily due to glycolysis. Agilent Seahorse XFp Glycolytic Rate Assay Kit User Guide...
- Page 8 Introduction Figure 2 Principle of the Agilent Seahorse XFp Glycolytic Rate Assay. Energy is produced by two pathways in the cell - glycolysis, and mitochondrial respiration. In the breakdown of glucose to lactate during glycolysis, protons are extruded into the extracellular media which is detected by the XFp Analyzer as ECAR.
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Page 9: Glossary
Introduction Glossary • Glycolysis: In the context of the Seahorse XFp Glycolytic Rate Assay, the process of converting glucose to lactate. • Buffer factor (BF): Buffer capacity of the measurement system, comprising the assay medium and XF assay conditions (instrument, sensor, labware). - Page 10 Introduction Agilent Seahorse XFp Glycolytic Rate Assay Kit User Guide...
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Page 11: Kit Contents
Rot/AA mix and 2-DG. The kit reagents are sufficient for six complete XFp Glycolytic Rate Assays in an Agilent Seahorse XFp Cell Culture Miniplate. Table 1 Agilent Seahorse XFp Glycolytic Rate Assay kit contents in each foil pouch ... -
Page 12: Additional Required Items
Kit Information Additional Required Items The following items are required to run the Seahorse XFp Glycolytic Rate Assays, but not supplied with the kit. Items Supplier Catalog number Agilent Seahorse XFe/XF Analyzers Agilent Technologies Seahourse XFp FluxPack (cartidges, 103022-100 Agilent Technologies miniplates, and calibrant) ... -
Page 13: Assay Workflow
Agilent Seahorse XFp Glycolytic Rate Assay Kit User Guide Assay Workflow Day Prior to Assay Day of Assay Running the Assay Analysis Using the Agilent Seahorse Glycolytic Rate Assay Report Generator Figure 3 Agilent Seahorse XFp Glycolytic Rate Assay workflow... -
Page 14: Day Prior To Assay
Day Prior to Assay For adherent cells, plate the cells at a predetermined density in the Agilent Seahorse XFp Cell Culture Miniplate using the appropriate cell culture growth medium. For more information, refer to the Basic Procedure, “Seeding Cells in Seahorse XF Cell Culture Microplates”, available on the... - Page 15 Warm the assay medium to 37 °C in a water bath. It is ready to use. Prepare Agilent Seahorse XFp cell culture miniplate for assay For adherent cells Remove the cell culture miniplate from the 37 °C CO incubator, and examine the cells under a microscope to confirm consistent plating and proper cell morphology.
- Page 16 Seahorse XFp Port A (Final well) Stock volume Medium 10X (Port) Rot/AA (µL) volume (µL) (µM) (µM) Port B (Final well) Stock volume Medium 10X (Port) 2-DG (µL) volume (µL) (mM) (mM) Agilent Seahorse XFp Glycolytic Rate Assay Kit User Guide...
- Page 17 Starting well assay medium volume and compound injection volumes Agilent Seahorse Starting well volume: XFp Analyzer 180 µL assay medium Port Vol. Conc. 20 µL 22 µL 25 µL 27 µL Agilent Seahorse XFp Glycolytic Rate Assay Kit User Guide...
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Page 18: Running The Assay
Continue. Calibration takes approximately 20 minutes. When calibration has been completed, remove the utility plate and place the Seahorse XFp Cell Culture Miniplate on the tray and click Continue to start the assay. Agilent Seahorse XFp Glycolytic Rate Assay Kit User Guide... -
Page 19: Analysis Using The Agilent Seahorse Glycolytic Rate Assay Report Generator
Analysis Using the Agilent Seahorse Glycolytic Rate Assay Report Generator When your run finishes, insert an USB Flash Drive on the front USB of the Seahorse XFp analyzer and save your assay file (*.asyr). In your Desktop version of Wave 2.3 or higher, open the file from your USB Flash Drive. - Page 20 Assay Workflow Agilent Seahorse XFp Glycolytic Rate Assay Kit User Guide...
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Page 21: Frequently Asked Questions
Agilent Seahorse XFp Glycolytic Rate Assay Kit User Guide Frequently Asked Questions Why does the assay require the use of buffered media, will it dampen the ECAR signal? A low concentration of HEPES (5 mM) has been found to provide consistent buffer capacity values across the time frame of the assay. - Page 22 ECAR values, to accurately calculate PER and glycoPER, phenol red must be omitted from the assay media. Questions: Contact Seahorse Technical Support: Email: Seahorse.support@agilent.com Ph: 800-227-9770, option 3, option 8 Agilent Seahorse XFp Glycolytic Rate Assay Kit User Guide...
- Page 24 Agilent Technologies For research use only. Not for use in diagnostic procedures. © Agilent Technologies, Inc. Printed in USA, May 2019 Revision B0 *103346-400* 103346-400...