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Agilent Technologies, Inc.

Seahorse XF T Cell Metabolic Profiling Kit

The Agilent Seahorse XF T Cell Metabolic Profiling kit allows for robust and accurate measurements of both glycolytic and mitochondrial activities in T cell populations, providing a complete picture of T cell energy metabolism. These measurements can be linked to antitumor properties of T cell therapy products and are therefore valuable in designing and optimizing therapy development processes to improve T cell persistence or avoid exhaustion in the tumor microenvironment. The kit not only provides improved reagents, but also features a streamlined assay workflow, reducing assay preparation time and minimizing the need for uncoupler optimization. The XF T Cell Metabolic Profiling kit is also integrated with Wave Pro and Seahorse Analytics software to simplify data analysis, visualization, and interpretation.

Each assay kit contains sufficient materials for six full-plate tests and is available in two packaging sizes. The XF kit is for use with Agilent Seahorse XF Pro and XFe/XF96 analyzers. The XFp kit is for use with the Agilent Seahorse HS Mini and XFp analyzers.

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  • Supports two assays for generating robust bioenergetic parameters linked to critical attributes for antitumor properties: T cell persistence and T cell metabolic fitness
  • Provide a comprehensive view of T cell energy metabolism
  • Suitable for evaluation of different construct designs, engineering strategies, starting material selection, or metabolic conditioning during the cell expansion process
  • Applicable for use in assessing the capacity of T cells to maintain metabolic fitness in metabolically restrictive conditions (e.g., tumor microenvironment)
  • Improved reagents to minimize optimization of uncouplers for different T cell populations
  • Validated for and best performed with Agilent Seahorse XF Pro analyzer, XF HS mini analyzer, XF RPMI medium, pH 7.4 (Cat. No. 103576-100) and supplements (Cat. No. 103577-100, 103578-100, 103579-100)
  • 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
  • Easy-to-interpret data analysis provided by the Agilent Seahorse Analytics, a cloud-based platform, simplifying data processing and transforming raw data into organized, shareable summary reports
  • Use this link to access Agilent Seahorse Analytics for data analysis: https://seahorseanalytics.agilent.com
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How it works

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T Cell Immunometabolism Fundamentals Revealed Through Seahorse XF Technology

The metabolic health of immune cells is a critical determinant that is intrinsically linked to antitumor efficacy, including performance within the immunosuppressive tumor microenvironment. Therefore, the analysis of immune cell metabolism offers unique insights into the critical drivers behind immune responses and provides opportunities to optimize cell therapy products.
Optimized for T cells, the new Agilent Seahorse T Cell Metabolic Profiling kit employs modulators with minimal cytotoxic effect, allowing for more accurate and robust measurements. This kit supports two assays.

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XF T Cell Persistence Assay

The T cell persistence assay is designed to investigate the ability of T cells to develop durable immune response and to generate memory-like T cells after infusion. Several publications indicate that distinct metabolic profiles of T cells (e.g., as a high spare respiratory capacity combined with a high oxidative and low glycolytic basal activities) are characteristics linked to improved T cell persistence after adoptive cell transfer. This assay can be used to evaluate of different construct designs, engineering strategies, starting material selection, or metabolic conditioning during the cell expansion process.

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XF T Cell Fitness Assay

The T cell fitness assay allows for the measurement of both T cell metabolism and fuel utilization under varied nutrient conditions, thus assessing the ability of T cells to sustain antitumor function in  metabolically restrictive environments, such as the ones encountered in the tumor microenvironment. Such conditions can be created by using substrate pathway inhibitors (e.g., etomoxir, UK5099, BPTES), replacing glucose by other carbon sources, or by directly altering levels of metabolic substrates in the assay medium.



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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