When should I use flow cytometry for signaling instead of western blot?


Posted by Chris S on Jan 9, 2019 3:15:00 AM

Flow cytometry enables quantitative analysis of protein expression, signaling states, and physical characteristics (cell size/granularity) at the single-cell level. Modern flow cytometers are capable of collecting data on multiple proteins from thousands of cells per second in a heterogeneous mixture. While flow cytometry is commonly employed to identify cell types using phenotypic markers expressed on the cell surface, it can also be used to measure intracellular signaling events.

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Researchers pursuing hypotheses about signaling events traditionally start with western blot. In this technique, lysates are pooled and analyzed one protein at a time, and infrequent signaling events may be missed due to insufficient detection sensitivity. Western blotting has limited capability to perform multiparameter quantitative analysis. Another drawback is that workflows can take a day or more to complete.

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Flow cytometry is an ideal solution to avoid these shortcomings, enabling multiplexed and quantitative analysis of signaling events with greater sensitivity and precision than western blot, in just a few hours. Importantly, by collecting discrete data points from individual cells, flow cytometry enables researchers to identify and quantify subpopulations of cells that exhibit specific signaling responses, to explore the range of responses in those cells, and to correlate multiple quantitative readouts. In short, signaling flow cytometry enables you to ask new research questions and characterize important signaling events from the plasma membrane to the nucleus.

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Get Started with Flow

We offer a diverse selection of antibodies and reagents for flow cytometry, supporting both quan­titative analysis of intracellular signaling pathways and phenotyping. Our expertise in antibody design, conju­gation, and flow cytometry enables us to develop and validate antibodies that you can use in your flow cytometry experiments with confidence. And the same scientists who develop and validate our flow cytom­etry products also provide technical support to ensure your experiments are successful.

Want to learn more? Download Our Signaling Flow Brochure

For Research Use Only. Not For Use In Diagnostic Procedures.

Topics: techniques, Flow

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