T-Cell Immunology
Technologies
Using panels of directly-conjugated fluorescent antibodies to recognize T cell specific markers, multicolor flow cytometric analysis allows researchers to interrogate specific target molecule levels expressed by individual cells in various phases of development.
Using flow cytometry and assay systems such as intracellular flow cytometry and BD™ Cytometric Bead Array (CBA) that measure proteins derived from activated T cell subsets, researchers have gained a detailed view of the cellular pathways and molecular mechanisms that support T cell development.
BD Biosciences also offers other complementary technologies including BD OptEIA™ ELISA reagents for individual cytokine quantitation, BD ELISPOT reagents for determining the frequency of cytokine-producing cells, and in vivo capture assays for quantitating the immune response by directly capturing cytokines in vivo to further advance research in the T cell area. Reagents and assays for intracellular staining are also available.
Intracellular Flow Cytometry
Intracellular flow cytometry has established its position as an extremely powerful tool for analysis of the nature and frequency of cells that produce cytokines, chemokines, inflammatory mediators, and signaling proteins. BD Biosciences offers several separate technologies for intracellular flow cytometry:
BD Cytofix/Cytoperm Methodology
The BD Cytofix/Cytoperm™ method and related reagents offers flexibility in intracellular cytokine staining. The protocol has been developed to work not only on human cells, but also in non-human primate, mouse, and rat samples. A wide range of antibodies and reagents that have been optimized for intracellular staining are available.
BD FastImmune
The BD FastImmune™ system is designed for intracellular cytokine detection by flow cytometry. It allows for quantitative and qualitative measurements of antigen-specific T cells in research studies to assess the importance of the monitoring of immune status during disease and in assessing vaccine efficacy. Intracellular staining has been made possible by the advent of high-affinity anti-cytokine antibodies, optimized cell fixation, and permeabilization protocols.
BD Phosflow Technology
BD™ Phosflow is a highly innovative flow cytometry–based technology, enabling activation state analysis of multiple proteins at the single-cell level. Using phosphorylation site-specific antibodies in combination with cell-surface markers, phosphorylation events can be studied directly in small subpopulations of complex primary samples.
Our proprietary buffer system also enables phospho-protein analysis directly in whole blood samples, minimizing artifacts caused by cell isolation steps and providing more relevant information as assays are performed at near-native conditions. This revolutionary technology enhances cell signaling network analysis and high-content secondary screening for drug discovery and development, and enables the identification of signaling pathway-dependent bio-signatures for disease.
BD Biosciences is developing an expanding line of phosphorylation site-specific antibodies for the study of intracellular protein phosphorylation using multicolor flow cytometry. BD Phosflow reagents are high-quality antibodies conjugated to a variety of fluorophores making multiparameter studies of phosphorylation events easier. Combine them with cell-surface immunophenotyping using our extensive selection of surface marker specificities to make an extremely powerful means of obtaining kinetic information on subset-specific signaling processes.
Technologies for Cytokine Measurement
BD Cytometric Bead Array (CBA)
BD CBA is a flow cytometry application that allows users to quantify multiple proteins simultaneously. The BD CBA portfolio includes assays for measurement of a variety of soluble and intracellular proteins including cytokines, chemokines, growth factors, and phosphorylated cell-signaling proteins. This technology makes CBA especially suitable for studying events involved in T cell differentiation.
Learn more about BD Cytometric Bead Array>BD™ ELISA Reagents
Whether relying on pre-optimized, ready-to-use ELISA systems, or developing custom assays, BD offers maximum flexibility. BD offers a large menu of optimized BD OptEIA™ kits and sets, and allows customization of ELISA systems by combining individual matched capture and detection antibody pairs with corresponding recombinant protein standards.
Learn more about BD ELISA reagents>BD™ ELISPOT Technology
The enzyme-linked immunospot (ELISPOT) assay is a powerful tool for detecting and analyzing individual cells that secrete a particular protein in vitro. This technology allows detection of cytokine releases at the single-cell level. It’s highly sensitive and allows for downstream analysis.
Learn more about BD ELISPOT technology>In Vivo Capture Cytokine Assays
This is a complimentary technology to ELISA and CBA that allows sensitive and direct measurement of cytokine production in vivo, in mice, through the injection of a cytokine-specific antibody and subsequent analysis of collected serum in an ELISA-type assay. The in vivo capture assays are easy-to-use and yield highly-sensitive and directly-correlated results.
Learn more about in vivo capture cytokine assays>