Flow Cytometry

Troubleshooting Guide: Fluorokine Flow Cytometry Kits

Fluorokine® Receptor Detection Kits are designed as alternative reagents for the detection of cell surface cytokine receptors by flow cytometry. Although the staining procedure of cells with this line of reagents is straight forward, some situations can present difficulties in data interpretation. In the table below, we attempt to point out some common problems that users of Fluorokine Receptor Detection Kits may encounter. The variable nature of cytometry instrumentation and instrument set-up can dramatically influence the quality of data generated with these reagents.

Flow Cytometry Protocol for Staining Intracellular Molecules using Detergents to Permeabilize the Cell Membrane

Flow cytometry can be used to analyze various intracellular molecules including phosphorylated signaling proteins and cytokines. Cytokines and other secreted molecules can be detected by flow cytometry in activated cells with the aid of secretion inhibitors, such as monensin or brefeldin A. These compounds prevent the export of newly synthesized proteins by disrupting the ER-Golgi transport machinery.

Flow Cytometry Protocol for Analysis of Cell Viability using Propidium Iodide

Flow cytometry provides a rapid and reliable method to quantify viable cells in a cell suspension. Determination of cell viability is critical when evaluating the response to cytotoxic drugs or other environmental factors. In addition, it is often necessary to detect dead cells in a cell suspension in order to exclude them from the analysis. Dead cells can generate artifacts as a result of nonspecific antibody binding or through unwanted uptake of fluorescent probes.

Analysis of Cell Viability using 7-Amino Actinomycin D (7-AAD)

Flow cytometry provides a rapid and reliable method to quantify viable cells in a cell suspension. Determination of cell viability is critical when evaluating the physiological state of cells, such as in response to cytotoxic drugs and environmental factors, or during the progression of cancer and other disease states. In addition, it is often necessary to detect dead cells in a cell suspension in order to exclude them from analysis.

Flow Cytometry Protocol for Staining Intracellular Molecules using Alcohol to Permeabilize the Cell Membrane

Flow cytometry can be used to analyze various intracellular molecules including phosphorylated signaling proteins and cytokines. Cytokines, small signaling molecules secreted by many cell types, can be detected by flow cytometry in activated cells with the aid of secretion inhibitors, such as monensin or brefeldin A. These compounds prevent the export of newly synthesized proteins by disrupting the ER-Golgi transport machinery.