I am carrying out Western blot to determine effects of extracts on expression of LPS induced iNOS in murine macrophage cells, I am using the iBind flex system from Thermofisher for multiplexing the membrane with Abs. One of the primary antibodies I am using is conjugated with a fluorochrome, and another is a unconjugated Ab. The Thermofisher manual for iBind system mentions about mixing the primary Abs. I am confused whether both the Primary Abs I am using should be used together to make up one mixture and added to the system or the conjugated primary Ab should be used along with the secondary Ab which is conjugates with another fluorochrome?