I've been trying the anti beta-actin loading control (Sigma) for Western blotting with Drosophila melanogaster protein extracts. The problem is that I tried lots of concentrations (1:250, 1:1500, 1:2500, 1:5000, 1:7500, 1:10000) and I didn't get any result. I used secondary antibody anti-mouse-POD at 1:5000, which works perfectly with the primary antibody I use to detect my protein of interest and with anti-tubulin loading control.
Can anybody help me, please? If someone had the same problem, a piece of advice will be much appreciated.
Thank you very much!