Intracellular staining and detection of cytokines in combination with cell surface antigen detection
Features
- Determination of immunological presentation of activated cells in disease-related immuno-monitoring
- Based on immuno-phenotype, but also on functional activity
- Developed for intracellular staining and detection of cytokines in combination with cell surface antigen detection
- Complete kit for fixation, permabilization and stimulation of the leukocytes to enhance the production of specific intracellular cytokines
- Each sample can be further analyzed for five different intracellular antigens using direct conjugated monoclonal antibodies
Clinical Applications
- Analysis of production of specific cytokines on single cell level
- Suitable method to monitor immune-(re)activation
- Detection of different subpopulations (like Th0 cells, Th1 cells and Th2 cells).
- Transplant patients
- Atopic disease
- HIV / AIDS patients
Research Applications
- Research on murine cells
- Detection of TGF-ß using our anti-TGF-ß antibody (IQP-169R)
Introduction
The Cytodetect™ Kit is intended for stimulation of leukocytes prior to analysis of intracellular antigens like cytokines and immunophenotyping of corresponding cells. Detection is performed using monoclonal antibodies selected for specific reactivity towards these antigens. Monoclonal antibodies are fluorochrome-conjugated for easier, direct application in flow cytometry analysis.
This procedure allows detection of different subpopulations of lymphocytes or leukocytes based on both immunophenotype and functional activity with respect to intracellular production of specific cytokines. Possibly, discrimination of different types of leukocytes based on their cytokine profile allows assignment of functional activity, as has been suggested for Th1 and Th2 cells. Distinction may be helpful in determination of immunological presentation of activated cells in disease-related immunomonitoring.
Principles of the Cytodetect™ Kit
The level of most of the cytokines produced by immune unstimulated cells is to low to be detected by flow cytometry analysis. Therefore, a method has been developed to analyze cells which have been stimulated before detection of these intracellularly expressed cytokines. The Cytodetect™ Kit provides reagents for cell stimulation, fixation and permeabilization, according to the sequential steps as listed below.
Three procedures are provided, a General Test Procedure which can be followed for immunostaining of e.g. CD3-positive or CD8-positive cells, an Alternative Test Procedure developed for immunostaining of CD4-positive cells and a slightly modified General Test Procedure for detection of TGF-ß. The Alternative Test Procedure results in a better discrimination of CD4-positive from CD4-negative cells, and thereby facilitating gate setting for flow cytometry analysis.
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