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Agilent Technologies, Inc.

Seahorse XF Hu T Cell Activation Assay Kit

Agilent Seahorse XF Hu T Cell Activation Assay Kit measures human (Hu) T cell activation response within several minutes of stimulation using Seahorse XF Analyzers. This is accomplished via measuring proton efflux rates (PER) associated with glycolytic pathway-dependent energy production, and is built on the principle that T cell activation is accompanied by a rapid switch in cellular energy production and intermediate demand to support effective proliferation, size expansion, and differentiation. Such principle is supported by recent advances in the field of immunology that recognize cellular metabolism as a primary driver and regulator of immune cell function and lineage commitment.

The kit contains Agilent-validated ImmunoCult Human CD3/CD28 T Cell Activator and 2-deoxy-D-glucose (2-DG) and is an optimal solution delivered by Agilent and STEMCELL Technologies partnership for interrogating and modulating T cell activation on the Seahorse XF platform. The ImmunoCult Human CD3/CD28 T Cell Activator from STEMCELL Technologies is a soluble tetrameric complex. It is easy-to-use and compatible with specially designed injection ports on Agilent Seahorse XF cartridges, reducing assay preparation time and improving assay reliability. Together with standardized protocols and cloud-based Agilent Seahorse Analytics software, the XF Hu T Cell Activation Assay Kit provides a robust measure of T cell activation with new insights.

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Features

  • Detects human T cell activation in minutes, providing an early window to investigate cellular and signaling requirements associated with T cell activation
  • Provides kinetic readout for continuous monitoring of human T cell activation and modulation
  • Allows dosing of cells with test compounds/modulators during assay through the built-in injection ports
  • The soluble ImmunoCult Human CD3/CD28 T Cell Activator (STEMCELL Technologies) is easy-to-use and compatible with specially-designed injection ports on Agilent Seahorse XF cartridges, reducing assay preparation time and improving assay reliability
  • Reagents are directly reconstituted or diluted in the assay medium, eliminating the need for DMSO or other solvents
  • Every lot is tested 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 T Cell Activation assay, share with collaborators, or export data to Microsoft Excel or GraphPad Prism for other graphing or analysis needs

How it works

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A rapid early increase in glycolysis as a signature event for T cell activation

Antigen presenting cell (APC) or CD3/CD28 activators, upon recognizing and binding to T cell receptors, initiate a number of signaling cascades that lead a rapid increase in glycolysis and subsequent T cell proliferation, cytokine production and effector functions. It is recognized that the rapid early onset of glycolysis is a signature event of T cell activation and is required for subsequent T cell functions.

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Seahorse XF technology offers a robust method for detecting T cell activation

The Agilent Seahorse XF Hu T Cell Activation Assay measures T cell activation response by monitoring glycolysis changes induced by activators. This rapid assay presents an early window to discriminate activation responses and provides kinetic functional information that typically cannot be obtained by orthogonal methods measuring gene expression, cell proliferation, or cytokine production. With standardized protocols and validated reagent, the XF Hu T Cell Activation Assay is a robust solution for studying T cell activation, modulation and related metabolic reprogramming.

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Easy-to-use turnkey solution

The XF T Cell Activation Assay is a turnkey solution with standardized protocols, ready-to-use XF PDL cell culture microplates and streamlined data analytic tool. The standardized protocols support two experimental designs that allow for testing/comparing activation potential among experimental groups and for investigating acute effects of modulators. Organized/shareable summary reports are automatically generated by Agilent Seahorse Analytics. Precoated XF PDL cell culture microplates provide convenience and redcuce data variation commonly caused by manual coating processes.

Resources

Downloads

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

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