endothelial cells and GPI-anchored proteins

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nikkie
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endothelial cells and GPI-anchored proteins

Post by nikkie » Mon Dec 11, 2006 7:42 am

As published you can purify GPI-anchored proteins (expressed in CHO cells) and add them exogenously to cells. The protein will incorporate into the cell membrane and can be detected onto the cell surface by FACS.
This is a standard method in our lab, and it works for nearly all cell types (tumour cells, CHOs, fibroblasts) The problem is that I cannot show this for primary endothelial cells (HUVECs, HDMECs), but for immortalized endothelial cells. Our idea is that the glycocalix is different.
The problem is that I really have to do this: so does anyone have an idea how to incorporate this GPI-anchored protein into primary endothelial cell membranes!!!

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