Application Examples
| A broad range of applications with the Cell Flourescence LabChip kit and cell assay extension include: |
Detection of fluorescent proteins
Detect the expression levels of various green fluorescent proteins (GFP)
A counterstain with a red live-cell stain serves to detect both – GFP-positive and GFP-negative cells. This assay is useful in monitoring the success of a wide range of transfection experiments during its optimization process.
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Detection of antibody binding
Detect the expression of non-fluorescent, cell surface and intracellular proteins by antibody staining.
Cells can be differentiated depending on the expression of a protein marker. The high sensitivity, laser based red channel is used to detect and quantify signals from an antibody stain like APC (allophycocyanin) or Cy5®. A counterstain with the blue live-cell stain calcein allows to gate on living cells.
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Detection of apoptotic cells
Analyze annexin V stained cells allowing detection of apoptotic cells early in the apoptotic process.
Apoptotic cells can also be detected by intracellular antibody staining of active caspase-3. Annexin V binds to phosphatidylserine (PS) – a membrane lipid that is kept to the inner leaflet of the cell membrane of intact cells. Exposure of PS to the outer leaflet is an early indicator of apoptotic pro-cesses. Annexin V binding is made detectable by Cy5 staining of the annexin V via biotin-streptavidin interaction. Calcein staining of the cells is used as a live control to distinguish living and apoptotic cells from dead cells.
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