Hi Ahmed,

Assuming you have a multichannel image which consists of an image of your bands of interest and an image of your total protein stain you can perform total protein normalisation easily and quickly using our Phoretix 1D software which you can get a free trial of here: https://totallab.com/products/phoretix-1d-free-trial/

We don't have a video demo of how to do this on our YouTube channel yet however the steps would be:

Automatically identify your bands > select normalisation > select total lane volume setting > select reference channel as your total protein stain channel

If you don't have that image, you can combine two images to produce a multichannel image within the software to perform your comparison.

Best wishes,

Steven

Hello, I found a video tutorial on ImageJ for western blot analysis. Hope it helps https://www.youtube.com/watch?v=u-u3G7JhIAo