We are trying to express a mammalian transmembrane protein on gram negative bacterial surface.

1. Our transformation was successful; we are trying to use flowcytometry to validate the expression of the protein on the surface of the bacteria.

What parameters are crucial, I am unable to segregate the bacterial population.

The peak does not shift significantly between the stained (green) and unstained (blue.

What threshold parameters should be modified for running bacteria in flowcytometry.

2. What other techniques can be used to validate the bacteria for surface expression of the target protein.

Thank you