Hello, I am a PhD student in the Dept. of Pharmaceutics, Trinity College Dublin, and I have a query regarding flow cytometry. I have never used this technique, and although I have access to a machine, there is no-one in my research group who has any practical experience either. So please excuse me if these questions seem very basic. I am trying to demonstrate the presence of the asialoglycoprotein receptor on the Hep G2 cell line. Although this is well established in the literature, for validation and characterisation purposes I would still like to make sure my cells have this receptor. I have a reference for labelling the receptor with a secondarily labelled antibody, but there is little detail on the actual protocol, and I have been unable to contact the corresponding author. I am hoping to use a FITC-labelled glycoprotein as my ligand, and my main concern is how to create a suspension of normally adherent cells, and whether the labelling/washing step should be carried out before or after the suspending stage. If anyone can help me, I would be very grateful. Anthony McMahon Dept. of Pharmaceutics School of Pharmacy Trinity College Dublin 2 Ireland Tel: 00353 1 608 2454 Fax: 00353 1 608 2783 Dept. of Pharmaceutics School of Pharmacy Trinity College Dublin 2 Ireland Tel: 00353 1 608 2454 Fax: 00353 1 608 2783
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