Wondering if anyone has observed this phenomena before. I'm testing a new molecule I made on some cells and when blotting for some proteins I'm interested in, the beta actin control looks like it disappears for the higher concentrations of the molecule. Now I thought it could have been an error on my part from loading, but the thing that absolutely perplexes me is that the same phenomena is happening across 4 different gels and two different cell lines. What are the odds that I made the same loading error for only the same drug concentration 4 times?
Has anyone heard of beta actin changing? I think I did all of the cell lysate/protein normalization calculations correctly. Of course I'll repeat the experiment again, but can beta actin really be affected that much? I'm seeing it almost disappear, only at the higher molecule concentrations. I'll try GAPDH, but what if the cell is so stressed that GAPDH will also be affected? Is there a stain I should do?