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Seahorse XF Mito Stress Test Kits

Delivering a reliable and comprehensive view of live-cell metabolism

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The Agilent Seahorse Mito Stress Test is the widely recognized, well accepted standard assay for assessing mitochondrial function. Multiple parameters are obtained in one assay including basal respiration, ATP-linked respiration, maximal and reserve capacities, as well as non-mitochondrial respiration. Each kit contains the material sufficient for six full plate tests. The XF kit is for use with 96- or 24-well Seahorse Analyzers. The XFp kit is for use with 8-well Seahorse Analyzers.|A new version of this kit, the Seahorse XF 3D Mito Stress Test kit, provides higher amounts of reagents in order to perform functional assays with 3D models such as tissues, organoids and small organisms.|For Research Use Only. Not for use in diagnostic procedures.
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103015-100

Seahorse XF Cell Mito Stress Test Kit. Each kit contains six (6) single-use pouches. Each pouch contains one (1) each of oligomycin, FCCP, and rotenone/antimycin A. For use with Seahorse XFe/XF96 and XF/XF24 analyzers.

  • Unit:

    1 Kit

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User Manuals
Seahorse XF Cell Mito Stress Test Kit User Guide (PDF)

View our in-depth user guide for the XF Cell Mito Stress Test Kit.

  • Publication Part Number: 103016-400
  • Created: 17 Dec 2024
  • 2 MB
  • PDF
Supporting Documents
Chinese (Simplified) (China) Complete (PDF)
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User Manuals
Seahorse XFp Cell Mito Stress Test Kit User Guide (PDF)

Learn how to setup and operate this standard assay for measuring mitochondrial function in cells.

  • Publication Part Number: 103011-400
  • Created: 17 Dec 2024
  • 2 MB
  • PDF
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Technical Overviews
Measuring Mitochondrial Function of Matrix-Embedded Organoids Using the Agilent Seahorse XF Flex Analyzer (PDF)

Procedures and guidelines for establishing and maintaining organoid cultures within the Seahorse XF Flex organoid microplates and performing XF assays

  • Publication Part Number: 5994-8742EN
  • Created: 10 June 2026
  • 2 MB
  • PDF
Community Search Results

How to manage negative value OCR and ECAR for Mitochondrial Stress Test and Glycolysis Stress Test

How to manage negative value OCR and ECAR for Mitochondrial Stress Test and Glycolysis Stress Test

Hi, I am working with data from the Seahorse XF Cell Mito Stress Test and Glycolysis Stress Test, and few wells show negative OCR or ECAR values at certain measurement points.

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BSA quantification using normalize function for a cell mito stress test

BSA quantification using normalize function for a cell mito stress test

I was hoping to use BSA quantification for normalization for a cell mito stress test. I am not sure if I should input my absorbance values or concentration values in the input fields, and what scale factor to use.

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Agilent Seahorse Mito Stress Test Single-File and Mulit-File XF Report Generator FAQs

Agilent Seahorse Mito Stress Test Single-File and Mulit-File XF Report Generator FAQs

Custom Cycles are not part of the standardized assay template for the Seahorse XF Cell Mito Stress Test and not supported in Report Generator analysis. Please contact Seahorse Technical Support if you have any additional questions regarding Custom Cycles.

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Seahorse Cell Mito Stress Medium

Seahorse Cell Mito Stress Medium

For preparing seahorse XF cell mito stress test assay media, the Agilent's protocol suggests adding 1 mL 100mM pyruvate solution (final pyruvate conc = 1mM), 1 mL 200 mM glutamine solution (final glutamine conc = 2 mM) and 1 mL 1.0 M glucose solution (final glucose conc= 10mM) in 97 mL XF base media.

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Applications
A Superior System for Real-Time Metabolic Analysis with Brain Tissue and Other 3D Models (PDF)

An optimized Agilent XF 3D tissue workflow that allows for detection and quantitative assessment of energy metabolism in tissue materials is presented.

  • Publication Part Number: 5994-8309EN
  • Created: 30 Apr 2026
  • 3 MB
  • PDF
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User Manuals
Agilent Seahorse XF 3D Mito Stress Test Organoid Workflow Quick Start Guide (PDF)

Agilent Seahorse XF 3D Mito Stress Test Organoid Workflow Quick Start Guide For use with the Agilent Seahorse XF Flex analyzer and the Agilent Seahorse XF Flex organoid microplate.

  • Publication Part Number: D0139619 Rev. A.01
  • Created: 13 Jan 2026
  • 347 KB
  • PDF
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User Manuals
Agilent Seahorse XF Mito Stress Test Islet Workflow Quick Start Guide (PDF)

Seahorse XF Mito Stress Test Islet Workflow Quick Start Guide for use with the Agilent Seahorse XF Flex analyzer and the Agilent Seahorse XF Flex 3D capture microplate-S.

  • Publication Part Number: D0138239 Rev A.01
  • Created: 13 Jan 2026
  • 641 KB
  • PDF
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Technical Overviews
Improving Real-Time Metabolic Analysis in Islets and Spheroids Using the Agilent Seahorse XF Flex 3D Capture Microplate-S (PDF)

In this technical overview, we present an optimized workflow for analyzing pancreatic islet metabolism with enhanced sensitivity, and reduced sample requirements using the Seahorse XF Flex analyzer and the Agilent Seahorse XF Flex 3D capture...

  • Publication Part Number: 5994-8739EN
  • Created: 31 Oct 2025
  • 3 MB
  • PDF
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User Manuals
Agilent Seahorse XF 3D Mito Stress Test Tissue Workflow Quick Start Guide (PDF)

Agilent Seahorse XF 3D Mito Stress Test Tissue workflow quick start guide for use with the Agilent Seahorse XF Flex Analyzer and the Agilent Seahorse XF Flex 3D Capture Microplate-L.

  • Publication Part Number: 5994-8268EN
  • Created: 28 Apr 2025
  • 433 KB
  • PDF
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Applications
Gaining Insights into Disease Biology for Target Identification and Validation (PDF)

This Application Note shows how Seahorse XF technology can be used to determine specific metabolic states associated with cellular dysfunction and disease

  • Publication Part Number: 5994-0838EN
  • Created: 19 Apr 2019
  • 573 KB
  • PDF
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Selection Guides
Seahorse XF Media and Buffer Selection Guide (PDF)

Seahorse XF media is specially formulated for use in Seahorse XF assays. This Selection guide lists media and buffer compatibility with the standard assays.

  • Publication Part Number: 5991-7878EN
  • Created: 11 Apr 2024
  • 1 MB
  • PDF
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Flyer
Agilent Seahorse XF Assay Kits and Reagents (PDF)

This documents contains high level description for all Agilent seahorse XF kits and reagents, include key features and benefits

  • Publication Part Number: 5991-8291EN
  • Created: 28 Apr 2025
  • 877 KB
  • PDF
Supporting Documents
Chinese (Simplified) (China) Complete (PDF)
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Agilent Cell Analysis Webinar Series

View informative Agilent Cell Analysis Technology webinars on a variety of research subjects.

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Posters
Real-Time Metabolic Profiling in Brain Tissue Biopsies Using the Seahorse XF Flex System (PDF)

Poster presented at SFN25, featuring the Agilent Seahorse XF Flex for real-time metabolic profiling in brain tissue biopsies.

  • Publication Part Number: Poster 107.05 (Board C19)
  • Created: 08 Dec 2025
  • 535 KB
  • PDF
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Selection Guides
Seahorse XF Instrument Selection Guide (PDF)

Instrument Selection Guide compares all XF Analyzer specifications

  • Publication Part Number: 5991-8215EN
  • Created: 25 Mar 2025
  • 461 KB
  • PDF
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Seahorse XF Glycolysis Stress Test Kit

Measure glycolytic capacity after glucose starvation

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The Agilent Seahorse Glycolysis Stress Test kit measures the capacity of the glycolytic pathway after glucose starvation. This is accomplished by driving cells toward glycolysis and assessing their ability to increase glycolytic activity to meet metabolic and bioenergetic demands. To measure cellular glycolysis rates under physiological conditions, please use the Agilent Seahorse Glycolytic Rate Assay.|The Glycolysis Stress Test kit contains the materials sufficient for six full plate tests and is available in two packaging sizes. Each kit contains the material sufficient for six full plate tests. The XF kit is for use with 96- or 24-well Seahorse Analyzers. The XFp kit is for use with 8-well Seahorse Analyzers.
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Seahorse XF Palmitate Oxidation Stress Test Kit and FAO Substrate

Seahorse XF Palmitate Oxidation Stress Test Kit includes reagents for 3 assays using XF Analyzers

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The Agilent Seahorse XF Palmitate Oxidation Stress Test Kit provides validated reagents to measure long chain fatty acid oxidation (FAO) via assessing changes in oxygen consumption (OCR) in intact cells. The kit includes the XF Palmitate-BSA FAO Substrate, L-carnitine, etomoxir, as well as oligomycin, FCCP, and rotenone/antimycin A to determine the intrinsic rate and capacity of a cell to oxidize palmitate in the absence or limitation of other exogenous substrates. This kit is best applied when investigating how interventions (genetic manipulations, drug exposure) specifically affect the long chain FAO process, and can be a complementary and/or follow assay to the XF Long Chain Fatty Acid Oxidation Stress Test. The kit is supplied with a standardized protocol to make it straightforward to perform the experiments and easy-to-use Seahorse Analytics to simplify data analysis.|The Seahorse XF Palmitate-BSA FAO Substrate included in the kit is pre-conjugated with a quantified free palmitate concentration eliminating the tedious, time-consuming, and variable conjugation process. The Palmitate-BSA reagent is also sold independently for use in other types of assays.
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Seahorse XF Mito Tox Assay Kit

Identify mitochondrial liability and de-risk your drug pipeline with Agilent XF Mito Tox solution

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The Agilent Seahorse XF Mito Tox Assay kit allows for the identification of drug-induced mitochondrial toxicity through direct functional measurements of mitochondrial oxygen consumption rates (OCR). This kit is adapted from the Agilent Seahorse XF Cell Mito Stress Test, the fundamental assay for interrogating mitochondrial function, with improved assay design and data outputs. It provides high sensitivity and specificity along with the toxicity mode of action in one assay, minimizing false positives and false negatives. The XF Mito Tox Assay Kit seamlessly integrates with Agilent Seahorse Pro Controller and Wave Pro software and Agilent Seahorse Analytics in the assay workflow, providing an end-to-end solution for assessing drug-induced mitochondrial toxicity. The software features greatly simplify the entire XF assay experience, from assay design or template creation to data interpretation, facilitating decision making to de-risk drug pipelines.|Each assay kit contains sufficient materials for six full-plate tests in 96-well format. The assay is only compatible with the Agilent Seahorse XF Pro analyzer.
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Seahorse XF Substrate Oxidation Stress Test Kits

Each Seahorse Substrate Oxidation Stress Test Kit provides reagents for 3 assays using XFe Analyzers

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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.

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