BD FastImmune Cytokine Detection System

Streamlined and highly reproducible intracellular cytokine staining for applied research applications on human samples

The BD FastImmune™ system is designed to meet the needs of applied research on human samples. Complete and optimized, the BD FastImmune system is particularly well suited for research studies monitoring immune status during disease, or immune responses to vaccine candidates.

BD Biosciences provides a range of BD FastImmune multicolor or single-color cytokine reagents for antibody staining, including selected sample-activation and sample-processing reagents.

Features

  • Whole blood method – Minimal sample manipulation and near in vivo environment
  • Streamlined protocol – Hands-on time of less than two hours, and results in as few as eight hours
  • Highly reproducible – Sufficient sensitivity to reproducibly detect rare, antigen-specific responses

BD FastImmune cytokine system workflow overview

Streamlined whole blood procedure

Developed for use with human whole blood samples, the BD FastImmune™ cytokine system procedure involves minimal sample manipulation and restimulation in close to in vivo conditions. The streamlined protocol involves a hands-on time of less than two hours and delivers results in as few as eight hours. Of particular importance for the analysis of antigen-specific activation, the BD FastImmune cytokine system provides sufficient sensitivity to reproducibly detect very low frequency responses, such as those of memory T cells that respond to specific soluble antigens in short-term restimulation assays.

BD FastImmune cytokine staining workflow

Several recent procedural developments add to the convenience when using this system in a pre-clinical workflow. To address the increasing need for standardization when using ICS assays in the pre-clinical setting, BD is involved in ongoing studies with the goal of optimizing inter-site data consistency.

Optimized reagents

BD FastImmune 3- and 4-color reagents are multicolor reagent combinations that have been optimized to work together and measure the most prevalent cytokine responses in either CD4 + T cells (IFN-γ, IL-2, TNF) or CD8 + T cells (IFN-γ). In addition to FITC-conjugated antibody to the cytokine, the 3- and 4-color reagents also include antibody to either CD4 or CD8 to set the acquisition gate, and PE Anti-CD69 to focus on the cells expressing this early activation antigen. Anti-CD4 and Anti-CD8 are conjugated to PerCP-Cy™5.5 for better separation of CD4 dim and CD8 dim T cells from the negative cell population. The 4-color formats also include CD3 staining to allow optimal gating on T lymphocytes (eg, discriminating from CD8 dim NK cells).

To make getting started easier, BD FastImmune detection kits also include a matching isotype control and necessary processing reagents, in addition to an optimized multicolor antibody reagent.

For added flexibility, all sample processing components and the multicolor reagents found in the kits are available individually. Additional single-color anti-cytokine reagents are also available for use with the BD FastImmune system, allowing researchers to extend analysis to cytokines not included in the preconfigured combinations.

Antigen-specific CD4+ IFN-γ responses

Detection of three cytokines simultaneously in CD4+ and CD8+ T cells

Cytokine expression in naïve, memory and effector T cells (part A)

Cytokine expression in naïve, memory and effector T cells (part B)


For Research Use Only. Not for use in diagnostic or therapeutic procedures.

Alexa Fluor® is a registered trademark of Life Technologies Corporation.

Cy™ is a trademark of Amersham Biosciences Corp. Cy dyes are subject to proprietary rights of Amersham Biosciences Corp and Carnegie Mellon University and are made and sold under license from Amersham Biosciences Corp only for research and in vitro diagnostic use. Any other use requires a commercial sublicense from Amersham Biosciences Corp, 800 Centennial Avenue, Piscataway, NJ 08855-1327, USA.

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