T Cell

Intracellular cytokine detection allows analysis of T-cell subset-specific responses

Detection of intracellular cytokines using multicolor flow cytometry is a powerful tool to gain insight into the spectrum of responses within mixed populations of cells. By simultaneously measuring multiple cytokines and cell surface markers in one assay, this technique offers insight into which cell subset is producing which cytokines. For example, multicolor flow cytometry can be used to identify Th1, Th2, or Th17 cells on the basis of their characteristic cytokine signatures. The cell-by-cell analysis allows researchers to analyze responses of distinct cell subsets without physically separating them.

Intracellular cytokine detection systems

For intracellular protein detection, cells must be fixed and permeabilized to allow a fluorescent antibody to enter and detect the target protein of interest. Different antigens have different sensitivities to and requirements for fixation and permeabilization, entailing additional optimization of protocols. To detect cytokines, which are secreted proteins, protein transport inhibitors are required to trap proteins inside the cells.

BD Biosciences offers two widely used assay systems for intracellular cytokine detection: BD Cytofix/Cytoperm™ reagents and the BD FastImmune™ cytokine system. Both systems offer researchers the ease and confidence of using optimized reagents and protocols and incorporate the high standard of quality and reproducibility that BD flow cytometry products are known for.

BD Cytofix/Cytoperm method

The BD Cytofix/Cytoperm method and related reagents address the wide-ranging experimental needs of basic research. With products for a broad selection of cytokines and multiple species, BD Cytofix/Cytoperm reagents offer maximum flexibility in intracellular cytokine staining.

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BD FastImmune system

The BD FastImmune system is designed to meet the needs of applied research on human samples. With a focus on complete, optimized systems, this approach is well suited for research studies monitoring immune status during disease, or immune responses to vaccine candidates.

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