I am incubating Herceptin with SKBR-3 cells to see the down regulation of HER2 receptor. I have incubated the cells for 4 hours and 24 hours, but they did not show any low expression, in fact they are showing higher expression (mean). Can anyone give some idea about how to get lower expression (which I found in all published papers)?
Thanks. Yes, I wanna see lowered HER2 levels upon Herceptin treatment with those cells which I found in lot of published papers. Actually I have used Herceptin (Trastuzumab) up to 500 ug incubated with MCF-7 (moderate expressed) and SKBR-3 (overexpressed) cells at two different time periods as per references (4 h and 24 h). After western blot and FACS analysis, I did not find any low HER2 expression or it's down regulation. Do you have any idea how I can get the expected result with any possible methods or ways?
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