I need to perform a western blotting using a N-Cadherin antibody. I used the antibody from BD Bioscences on hela, huvec and human fibroblasts, hoping these cells were good positive controls but I had no results. Do You know if the matter is caused by bad antibody supplied from BD or bad cells as positive control?
I guess I’d blame the cells before I’d blame the kit, but I don’t know enough about either to offer much advice. Make sure you’re diluting the antibody and doing any blocking correctly–but I imagine you’ve checked over obvious things like that. Maybe try a cell line of neurological origin?
Thank you for your advices, but unfortunatly I don’t have a cell line of neurological origin because I work with normal human keratinocytes that I have infected with a gene responsable of EMT. The only cell lines that I have are the ones that I have tested.