Flow Cytometry Animation


Cell processes are controlled by complex and
dynamic intracellular signaling networks. Surface receptors pass information into the
cell by modifying cytoplasmic proteins. These modifications include phosphorylation,. methylation, acetylation, which control a
broad range of cellular functions, including growth, development, and apoptosis. Disruption of signaling networks often leads
to disease. To understand signaling processes and normal
and pathological conditions, Identification and quantification of signaling proteins is
necessary. Flow cytometry is ideal for such a task. Flow cytometry is a laser based technology
widely used to phenotype cells using antibodies directed against surface markers. It can also allow simultaneous analysis of
multiple intracellular targets using antibodies that detects and signaling proteins, their
post-translational modifications, epigenetic modifications, and downstream transcription
factors. The first step of intracellular flow cytometry
is fixation of the cells using formaldehyde, which immobilizes proteins and preserves transient
signaling events. Cells are then permeabilized with methanol
or detergent, allowing antibodies to access to the intracellular space. Optimal fixation and premium realization conditions
depend on the location and biochemistry of the target epitopes. It shouldn’t be carefully considered to ensure
successful antigen detection, especially when staining surface and intracellular proteins
simultaneously. Since multiple signaling proteins and surface
antigens can be identified using fluorescently distinct antibodies, flow cytometry can readily
characterize signaling profiles at the single cell level within phenotypically distinct
populations. This allows the characterization of the relationship
between intracellular targets and enables detection of signaling events from rare cell
populations within a heterogeneous sample. Multiplex analysis of these protein targets
saves time and resources while producing data-rich results, making interests it flow cytometry
and invaluable tool to unravel the complexities of cell signaling for more information on
validated products and optimized protocols for flow cytometry, visit www.cellsignal.com/flowcytometry.

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