How to normalize a protein band to Housekeeping gene band using Biorad Image lab software?

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Yomna S. Momen

For more clarification, I have 2 membranes one for the B-actin and one for the protein of interest. For each membrane I have 8 lanes with 8 bands for the protein of interest, meaning that for each band of protein of interest I have a B-actin band.

Jack Chen

There may be variations even if you load to gels and transfer exactly the same.

Ideally, you should normalize your protein band of interest to the B-actin bands of the same lane on the same membrane. Then you can do the comparisons.

Thank You Jack. I appreciate your answer. Could I ask you if you use biorad image software for normalization and how? Thank you