Hi all,

I am currently working on Western Blots and trying to detect intact and cleaved proteins. To develop my membranes I am using the Chemidoc System from BioRad.

The point is that when I develop my membranes using the digital system, the strong Pro-Caspase-3 band is bleaching the weaker cleaved caspase-3 band (the antibody detects both) so the image appears as there is not cleaved caspase in the assay. I confirmed this hypothesis developing the same membrane in the old school (with films) and the cleaved band showed up.

After playing around with the software, I figure out that you can set up the sensitivity of the CCD Sensor the way you want (attached picture) and then have a good image that fits both bands.

I would like to know if anyone tried to find out which values for sensitivity matches the sensitivity of the Hyperfilm or Regular Kodak film?

Thank you a lot!