Antigen-specific T cells

Stimulation, magnetic enrichment, and flow analysis of antigen-specific T cells

  • Potent stimulation with PepTivator® Peptide Pools
  • Efficient enrichment of antigen-activated T cells
  • Reliable multiparameter flow analysis of antigen-specific T cells

Application protocols

Discover our different workflows for antigen-specific T cells and find the one that fits your experimental needs.

Application data by workflow step

PBMC isolation

Comparison of PBMC populations isolated with the MACSprep PBMC Isolation Kit or by density gradient centrifugation
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Fast PBMC isolation with the MACSprep™ PBMC Isolation Kit, human

  • Untouched isolation of PBMCs from 1–10 mL of human whole blood

  • No density centrifugation, no erythrocyte lysis

  • Fast processing in only 25 minutes

  • Superior reproducibility, recovery, and cell viability

T cell stimulation

Production of IFN-γ by human CD4+ or CD8+ T cells after stimulation with different antigens using PepTivator Peptide Pools
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PepTivator® Peptide Pools

  • Stimulation of both CD4+ and CD8+ T cells with our extensive panel of antigens

  • 15-mer peptides with 11-amino acid overlap that cover the complete sequence of the respective antigen

  • Available in 96-well format

  • MACS GMP-grade peptide pools for translational settings

Enrichment of antigen-specific T cells

Enrichment of antigen-specific CD4+ T cells using the CD154 MicroBead Kit, human
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Isolation of antigen-specific CD4+ T cells with the CD154 MicroBead Kit, human

The CD154 antibody specifically recognizes the human CD154 antigen, also known as CD40L, gp39, T-BAM, TRAP, or Ly-62. CD154 is transiently up-regulated on activated CD4+ T cells and plays an important role as a co-stimulatory molecule in T cell/antigen-presenting cell (APC) interactions through ligation of CD40. Due to its transient expression within hours after activation, CD154 can be used as a marker for isolation of activated antigen-specific CD4+ T cells. Optionally, pre-enrichment can be performed using the CD4+ T Cell Isolation Kit, human.

Isolation of antigen-specific CD8+ T cells with the CD137 MicroBead Kit, human

CD137 is mainly expressed on activated CD4+ and CD8+ T cells, activated B cells, and on natural killer cells, but can also be found on resting monocytes and dendritic cells. CD137 has been described to be a suitable marker for antigen-specific activation of human CD8+ T cells, as CD137 is not expressed on resting CD8+ T cells and its expression is reliably induced after 24 hours of stimulation.  For the isolation of activated antigen-specific CD8+ T cells, the CD137 MicroBead Kit can be combined with the CD8+ T Cell Isolation Kit.

Principle of the MACS Cytokine Secretion Assays
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Cytokine Secretion Assay – Cell Enrichment and Detection Kits

  • Unique technology for the isolation of viable cytokine-secreting cells

  • Accurate analysis of cytokine secretion at single-cell level

  • Sensitive detection of one T cell per million cells

  • Available for human and mouse cells secreting TNF-a, GMCSF, IFN-a, IFN-g, or IL-2, -4, -5, -10, -12, -13, -17, -22

Rapid ARTE – Enrichment of rare antigen-specific CD154+CD4+ T cells for sensitive flow analysis

By combining magnetic cell enrichment and multiparameter flow cytometry analysis of CD154+CD4+ T cells, Rapid ARTE allows the processing of larger cell numbers and the direct ex vivo detection and characterization of rare antigen-specific T cells.

  • Direct ex vivo detection and characterization of rare antigen-specific T cells

  • Fast protocol with only 1 hour hands-on time

  • Unique combination of cell enrichment via CD154 marker and multiparameter flow analysis

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Scientific poster
The rapid antigen-reactive T cell enrichment procedure enables direct ex vivo BKV-specific CD4+ T cell characterization

Michaela Niemöller, Mario Assenmacher, and Anne Richter
Miltenyi Biotec GmbH, Bergisch Gladbach, Germany

Antigen-specific T cell isolation for adoptive T cell therapy

The CliniMACS® Cytokine Capture System (IFN-gamma) allows the direct isolation of antigen-specific CD4+ and CD8+ T cells following a short in vitro stimulation period with a specific antigen. 

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Automatic isolation of antigen-specific T cells for adoptive T cell therapy

CD4+ and CD8+ T cells stimulated in vitro with a specific antigen can be magnetically labeled with the CliniMACS Cytokine Capture System (IFN-gamma). In combination with the new CliniMACS Prodigy®, the system allows fast and fully automated generation of antigen-specific T cells in a closed sterile system.

Flow cytometric characterization of antigen-specific T cells

As the final step in many workflows, flow analysis plays an important part in various T cell applications. We provide reagents and detailed application notes that help you make sure your flow analysis and phenotypic characterization of rare antigen-specific T cells are reliable and consistent.

  • REAfinity Recombinant Antibodies recombinantly generated antibodies that ensure lot-to-lot consistency and eliminate background signal
  • Rapid Cytokine Inspector – combined surface marker and intra-cellular cytokine staining for multiparameter analysis of activated human T cells
  • Cytokine Secretion Assay – Detection Kits – flow analysis of cytokine secretion at a detection level of one T cell in a million
  • Build your own T cell flow panel with our Flow Panel Builder
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Application note
Detection, enrichment, and direct phenotypic and functional characterization of antigen-specific naive and memory T cell subsets

Petra Bacher and Alexander Scheffold
Department of Cellular Immunology, Clinic for Rheumatology and Clinical Immunology, Charité, University Medicine Berlin, Berlin, Germany


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