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Shedding Light on Novel Cellular Pathways with TTP LabTech

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TTP LabTech has highlighted the use of the Acumen® eX3 laser scanning imaging cytometer to elucidate novel cellular pathways involved in the shedding of the epidermal growth factor receptor (EGFR).

The application note entitled “The use of Acumen eX3 for the identification of the roles of Protein Kinase C (PKC) α & δ isoforms in PMA-induced ectodomain shedding” describes how the high throughput imaging technology of the Acumen eX3 accurately and sensitively quantifies and monitors protein expression levels.

Online image capture and analysis, as well as multiplexing capabilities, allows the Acumen eX3 platform to provide consistent and accurate data from complex biological systems.

The shedding of HB-EGF is classically controlled by a disintegrin and metalloproteinase, ADAM17. The imaging technology of Acumen eX3 was used to determine the contributions of additional enzymatic pathways in this process.

HT1080 cells, transfected with a modified, alkaline phosphatase linked, heparin binding EGFR (AP-HB-EGF) were examined by both the traditional enzymatic assay and immunofluorescence for receptor shedding in the presence and absence of inhibitors of ADAM17 and PKC isoforms.

Experimental data suggest that important contributions are made by the PKC isoforms α & δ. Disruption of EGFR homeostasis is associated with a number of important cancer targets, notably breast, lung and colon tumours. Consequently research of this type is vital to further the fight against these diseases.