Abstract
Analysis of intracellular cytokines is extremely important in the clinical treatment of numerous diseases. Flow cytometry (FCM) is a highly effective technique that detects intracellular cytokines using specific fluorescence-labeled antibodies. The common steps of this assay include cell collection, fixation, permeabilization, blocking, intracellular staining and analysis by FCM. This technique also allows for analyzing the biological function of cytokines. In this chapter, we describe a modified method to detect the specific intracellular cytokine staining using FCM, with an emphasis on the effects of variables including samples, temperature, buffers, data acquisition, and analysis.
Publication types
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Research Support, N.I.H., Extramural
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Research Support, Non-U.S. Gov't
MeSH terms
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ATP Binding Cassette Transporter, Subfamily B, Member 1 / analysis*
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ATP Binding Cassette Transporter, Subfamily B, Member 1 / genetics
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ATP Binding Cassette Transporter, Subfamily B, Member 1 / metabolism
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Antibodies / chemistry
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Cell Line, Tumor
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Epithelial Cells / cytology
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Epithelial Cells / drug effects
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Epithelial Cells / metabolism*
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Flow Cytometry
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Gene Expression
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Humans
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Interleukin-8 / analysis*
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Interleukin-8 / genetics
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Interleukin-8 / metabolism
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Monensin / pharmacology
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Proton Ionophores / pharmacology
Substances
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ATP Binding Cassette Transporter, Subfamily B, Member 1
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Antibodies
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Interleukin-8
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Proton Ionophores
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Monensin