Cytoplasmic Protein Expression with Flow Cytometry

Using Flow Cytometry, an investigator can study protein expression in cells. Surface proteins may be detected using antigen-specific antibodies. Antibodies tagged with a fluorichrome are detected by the Flow cytometer. The RIC Facility FacsCanto can observe up to six fluorichromes in addition to size and granularity readings for each cell.

Fluorescent reporter proteins such as green fluorescent or red fluorescent proteins in transgenic, transfected, or transformed cells can also be detected. The Flow cytometer detects and reports the intensity of the fluorescent protein or fluorichrome-tagged antibody.

The above is an example of Flow data generated by analysis of a blood sample from a transgenic mouse expressing low and high levels of green fluorescent protein (GFP).

Non-fluorescent cytoplasmic proteins may also be observed by staining permeablized cells. The stain can either be a fluorescence compound able to bind to certain molecules, or a fluorichrome-tagged antibody to bind the molecule of choice.

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