I will bring bacteria and receptor binding protein together, protein binds bacteria.
Is there any differences between absorbances of just bacteria and after interaction of bacteria+receptor binding protein at Od600?
Or Bradford measurement of just receptor binding protein and bacteria+receptor binding protein ?
Thanks
It is good that you try to look at protein interaction using absorption spectroscopy. But it seems a very difficult to get the higher order interaction signal as compared to the scattering due to bacterial suspension as well as you can't specifically say that interaction is happening at desired site.
You have to do some interaction specific experiments using fluorescence spectroscopy i.e. using fluorescent probe labelled protein, FRET and anisotropy measurement.
Here is some link of research article may help you
https://ijpsr.com/bft-article/different-approaches-for-studying-protein-protein-interaction-and-analysis/?view=fulltext
Article Techniques for the Analysis of Protein-Protein Interactions in Vivo
Article Detecting ligand interactions in real time on living bacterial cells
thanks
The two approaches mentioned by you, viz., measuring OD@600 nm and Bradford measurement are very non-specific and it would not reveal the differences between a 'bacteria' & 'bacteria + receptor binding protein' what you are looking for!
If you want to see if the protein is binding to the bacteria, an easy way would be to use the bacteria to pull down the protein from solution. Mix the bacteria and protein together, incubate for a little while, spin down the bacteria, and measure the loss of the protein from the supernatant.
Another method would be to attach a fluorescent label to the protein, mix it with the bacteria, then look at the bacteria with a fluorescence microscope, or by flow cytometry.
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