Cytokine Secretion Detection
Cytokine Secretion Detection - Related Videos
11:53
ELISPOT Assay: Detection of IFN-γ Secreting Splenocytes
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2023
Source: Tonya J. Webb1 1 Department of Microbiology and Immunology, University of Maryland School of Medicine and the Marlene and Stewart Greenebaum Comprehensive Cancer Center, Baltimore, Maryland 21201 ELISPOT is a standardized, reproducible assay used to detect cellular immune responses. The assay utilizes an enzyme-linked immunosorbent assay (ELISA)- based method to detect single-cell immune responses which can be visualized by spots, hence the name ELISPOT. ELISPOT was first described in...
12:58
Expression of Exogenous Cytokine in Patient-derived Xenografts via Injection with a Cytokine-transduced Stromal Cell Line
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Cited by 2 •
2017
Described here is a method for producing exogenous cytokine in patient-derived xenograft (PDX) mice via weekly intraperitoneal injection of a cytokine-transduced stromal cell line. This method broadens the utility of PDX and provides the option for transient or sustained exogenous cytokine delivery in a multitude of PDX models.
09:43
Microfluidic Approach to Resolve Simultaneous and Sequential Cytokine Secretion of Individual Polyfunctional Cells
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Cited by 5 •
2024
The protocol describes an advanced microfluidic platform to quantitatively measure cytokine secretion dynamics of individual human peripheral blood mononuclear cells. The platform measures up to three cytokines in parallel (IL-6, TNFα, and IL-1β) for each individual cell stimulated with lipopolysaccharide as an example.
08:32
Dissecting Innate Immune Signaling in Viral Evasion of Cytokine Production
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Cited by 2 •
2014
We describe a protocol to measure the antiviral cytokine production in mice infected with a model herpesvirus, murine gamma herpesvirus 68 (γHV68) that is closely-related to human Kaposi’s sarcoma-associated herpesvirus (KSHV) and Epstein-Barr virus (EBV). Utilizing genetically modified mouse strains and mouse embryonic fibroblasts (MEFs), we assessed the antiviral cytokine production both in vivo and ex vivo. “Reconstituting” the expression of innate immune components in knockout embryonic...
05:00
Localized Surface Plasmon Resonance Imaging to Detect Protein Secretions from a Single Cell
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2025
This video demonstrates localized surface plasmon resonance imaging to detect protein secretions from a single cell. Using a microfluidic setup and a peptide-bound plasmonic nanostructure chip, changes in reflected light intensity at the resonance angle reveal interactions with secreted proteins from a single cell.
FAQs
The assay uses two antibody-based recognition steps: a capture antibody retains the cytokine, and a labeled detection antibody binds the captured protein. This interaction produces a measurable signal proportional to cytokine abundance in the tested material. The design converts secretion into a quantitative readout that supports comparisons among immune or inflammatory responses.
These formats answer different biological questions. ELISA measures cytokine abundance in culture fluid, whereas ELISpot identifies cytokine-producing cells. Multiplex bead assays measure multiple cytokines in one sample, enabling broader profiling. Selection depends on whether the priority is bulk secretion, the presence of producing cells, or simultaneous measurement of several signaling proteins.
A culture-fluid measurement reports the amount of cytokine present in the sample, while ELISpot identifies cytokine-producing cells. These readouts are complementary rather than interchangeable. Together, they can distinguish a strong overall secretion response from one generated by a particular cell population, improving interpretation of immune-cell activity.
Cells are first stimulated under controlled conditions, after which the relevant culture material or cell-based assay format is examined. For antibody-based measurement, the cytokine binds a capture antibody, a labeled detection antibody is added, and the resulting signal is measured. The signal indicates cytokine abundance or secretion according to the selected assay design.
It is useful when a study needs evidence of immune activation, cellular communication, or inflammatory responses. Measurements can compare cellular behavior under different experimental conditions and assess effects associated with vaccines, drugs, or disease-related states. The resulting secretion profile provides functional information about how immune cells respond.
In biology research, the method links molecular signaling with functional cell behavior. Cytokines provide measurable evidence of communication and inflammatory activity, so secretion patterns help characterize immune-cell function. Culture-fluid assays, cell-focused formats, and multiplex measurements offer complementary context for interpreting responses to stimulation, treatment, or disease-related conditions.