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I tested the antibody on human ovarian cancer cell extracts and A549 cell line. Samples were collected, sonicated, mixed with loading buffer, and resolved on a vertical electrophoresis system using denaturating conditions. I incubated the nitrocellulose membrane using 1:1000 antibody diluted in 5% non-fat milk with TBS-T for 2 h on rt, washed in TBS-T and incubated with adequate secondary antibody. Visualization was done using ECL. Signals were strong, but there was three of them, between the…
