header background-block header background-block header background-block header background-block

Find your product

Agilent Technologies, Inc.

Seahorse XF Substrate Oxidation Stress Test Kits

The Agilent Seahorse XF Substrate Oxidation Stress Test Kits are a suite of optimized solutions for rapidly measuring cellular substrate oxidation by assessing changes in oxygen consumption (OCR) in live cells. These kits allow for the interrogation of three primary substrates that fuel the mitochondria: long chain fatty acids , glucose/pyruvate, and/or glutamine, facilitating investigation of how cells alter or shift oxidation of specific substrates to perform key cellular functions of activation, proliferation, and differentiation, as well as to respond to genetic or pharmaceutical interventions or survival in microenvironments. By combining the well-recognized XF assay for mitochondrial function (Cell Mito Stress Test) with substrate pathway specific inhibitors (etomoxir, UK5099, BPTES), these kits report multiple functional parameters, providing rich and robust data. The kits are supplied with validated reagents, standardized protocols, and easy-to-use Seahorse Analytics to enhance the user experience.

filler image
filler image
filler image

Features

  • Discover how cell phenotype or fate can be controlled via manipulation or control of substrate oxidation activities
  • Understand which substrates are relevant or required for specific cellular function and/or phenotype using specific, optimized XF Substrate Oxidation Stress Test kits and protocols
  • Build on the well-accepted XF assay (XF Mito Stress Test) for interrogating mitochondrial metabolism: dive deeper into effects of specific substrate oxidation on cellular function, including activation, proliferation, differentiation, and dysfunction
  • Use specific substrate pathway inhibitors to reveal cells’ dependency on three primary mitochondrial substrates: long chain fatty acids, glucose/pyruvate, and/or glutamine under both basal respiration and high energy demand condition
  • May be used individually or in combination, allowing investigation of single or multiple substrates at the same time
  • Provides rich real-time multiparametric information not available from conventional endpoint substrate uptake or utilization assays
  • Compounds are reconstituted directly in the assay medium, eliminating the need for DMSO or other solvents
  • Every lot is rigorously tested and validated with functional assays to deliver consistent and robust results
  • Upload your assay result file to your Agilent Seahorse Analytics account to calculate and report parameters of the XF Substrate Oxidation Stress Test, share with collaborators, or export data to Microsoft Excel or GraphPad Prism for other graphing or analysis needs

How it works

filler image

Rapid Seahorse XF methods that measure mitochondrial substrate oxidation

Long-chain fatty acids (LCFAs), glucose/pyruvate and glutamine are three primary substrates that fuel mitochondrial metabolism. Researchers in many areas (e.g., cancer, immunology, stem cell biology) have demonstrated how oxidization of these substrates has profound impact on cell fate, function, and fitness. Many of the findings not only provide new biological insights, but also reveal novel approaches to intervention and developing successful research therapies. The Seahorse XF Substrate Oxidation Stress Test Kits enable rapid assessment of substrate oxidation in live cells.

filler image

Revealing substrate dependency under basal and high energy demand states

The assay involves an acute injection of a single pathway inhibitor (etomoxir, UK5099, or BPTES),  followed by 3 sequential injections of oligomycin, FCCP, and rotenone/antimycin A. This design provides information for basal respiration and acute response to the inhibitor (i.e., basal response), as well as maximal respiration, +/- the inhibitor. While minimal changes may be observed under basal conditions, larger responses are revealed in maximal respiration under high energy demand, which increases the dynamic window to measure changes in oxidation of specific substrates.

filler image

Flexible assay design and more kit options to address different research needs

The kits can be used individually or in combination, allowing testing of a single substrate or multiple substrates in one experiment. The basic procedure is performed in standard assay conditions and is best used for examining how mitochondrial substrate demand and reliance is affected when an intervention (e.g., a genetic manipulation or drug exposure) is applied to the cells.  
Note that an advanced protocol and a kit designed for explicit investigation of long-chain FAO using palmitate-BSA are also available. For details, visit the Agilent website for product 103693-100.

Resources

Downloads

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

Supplier

Agilent Technologies, Inc.

Visit Suppliers Website

Request a demo or a quote

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

Measure mitochondrial respiration, glycolysis, and ATP production in live cells in a 96-well format

Overview of Consumables for XF Analyzers

Simple, Automated, Charge-based DNA and RNA Extraction

LUNA-FX7™ Automated Fluorescence Cell Counter with unmatched cell counting accuracy

High multiplexing, high resolution in situ platform to combine single-cell and spatial genomics.

The best super-resolution microscopy experience, on your desktop.

NovoCyte Opteon Spectral Flow Cytometer Systems 3-5 Lasers and 73 detectors

The NovoCyte Penteon™ is a 5 laser benchtop flow cytometer with up to 30 colors

Benchtop Microfluidic Cell Sorting with Two Lasers and up to Nine Colors

An incubator designed specifically for the cultivation of primary human cells

Seahorse XFe24 Analyzers measure OCR and ECAR of live cells in a 24-well plate format.

Automated high content image acquisition and analysis for drug discovery and cell biology

Multicolor fluorescence imaging to data analysis in one device

IsoLight – Automated Cellular Proteomics Hub

IsoLight and IsoSpark – Single-Cell Functional Proteomics Hub

The LUNA-FL™ can count in both brightfield and fluorescence mode

The LUNA-II™ automated cell counter gives you the ultimate brightfield cell counting experience.

The MARS® System is the “one-stop” modular solution for complete cell isolation from complex samples

The NovoCyte Quanteon™ is a 4 laser benchtop flow cytometer with up to 25 colors

The QUANTOM Tx Microbial Cell Counter automatically identifies and counts individual bacterial cells

Single Cells or Nuclei from Solid Tissues in Minutes

Tapestri® Platform: Untangle the complexity of cancer with true multi-omics.

Sterile, easy-to-use solutions for high cell integrity of single cell dispensing

Tissue Clearing System for electrophoretic tissue clearing

Optimized Microplate Solutions for Research & Drug Discovery

DeepLabel™ Antibody Staining Kit for Cleared Tissues

IsoPlexis Solutions and Kits

Autofocusing cell counting and declustering algorithm that counts individual yeast cells

Automated Fluorescence Cell Counter for Stem Cells & SVF

The NovoCyte Advanteon provides the flexibility of 1, 2, or 3 laser options.

Accurate & efficient single-cell detection of SNVs, indels, CNVs and LOH across targeted gene panels

Measure functional metabolism in 3 dimensions from in vitro spheroids

NovoCyte Consumables and Accessories for your Flow Cytometry needs.

Kits and reagents for real-time fluorescence plate reader-based live cell metabolism assays

Sterile, easy-to-use solutions for high cell integrity of single cell dispensing

The best value benchtop flow cytometer with high flexibility, enhanced sensitivity and resolution.

Optimized solution for assessing mitochondrial function.

Provides real-time measurements of glycolysis

Measures human (Hu) T cell activation response within several minutes of stimulation

Formulated media based on common culture media DMEM or RPMI, and optimized for use in XF assays.

Analytical method for measuring basal state mitochondrial fuel oxidation in live cells

Identification of drug-induced mitochondrial toxicity

Provides validated reagents to measure long chain fatty acid oxidation (FAO)

For in vitro studies of long chain fatty acid oxidation by intact cells.

Permeabilizes intact cells in culture, selectively targeting the cellular plasma membrane.

Seahorse XF Real-Time ATP Rate Assay

A suite of optimized solutions for rapidly measuring cellular substrate oxidation

Robust & accurate measurements of both glycolytic and mitochondrial activities in T cell populations

An Error occurred please try again later.
Please fill in all mandatory fields.

* All mandatory fields must be completed.

Thank you for contacting us!

{{ successMessage }}