Pe mouse anti human her 2 neu

Flow cytometry analysis allowed measuring the expression of Her2 on the surface of PC-3 cells [28 (link)].
As previously described [29 (link)], QuantiBRITE PE (BD Biosciences, San Jose, USA) was used to estimate the absolute number of Her2 receptors on cell membranes. About 100,000 cells of PC-3 were incubated under saturated conditions with PE Mouse Anti-Human HER-2/neu (BD Biosciences, San Jose, USA) for 30 min at 4 °C before being rinsed with PBS. IgG2a-PE anti-mouse antibodies (Fisher Scientific, Illkirch, France) were used for isotopic control. Analysis was then immediately performed on Gallios Beckman Coulter. Assuming our anti-HER-2 PE antibody has a 1:1 fluorochrome/antibody ratio, PE median fluorescence intensity (MFI) was measured for all cell lines and reported on a log–log graph with MFI vs. PE molecules, after subtracting isotopic control MFI. All the measures were performed in triplicate.

1,2-distearoyl-sn-glycero-3-phosphoethanolamine-N-(aleimide(polyethyleneglycol)-2000) (Mal-PEG) and 1,2-dioleoyl-3-trimethylammoniumpropane (DOTAP) were purchased from COGER (Paris, France). Egg yolk phosphatidylcholine (PC) and cholesterol (Chol) were purchased from Sigma (St-Quentin-Fallavier, France). ASO was purchased from Eurofins (Les Ullis, France). 2-iminothiolane (Traut’s reagent) and Draq5 were purchased from Fisher Scientific (Illkirch-Graffenstaden, France). QuantiBRITE phycoerythrin (PE) and PE Mouse Anti-Human Her-2/neu were purchased from BD Biosciences (San Jose, CA, USA). Trastuzumab (Herceptin) was kindly given by Genentech (South San Francisco, CA, USA). All other reagents were of analytical grade.