Hi, I have been developing a BONCAT protocol for use on environmental marine bacterial samples using L-azidohomoalanine (AHA). From my testing and literature research, I found that incubating seawater samples for 3 hours with an AHA concentration of 1mM to produce the best results. However, when performing strain promoted click chemistry with the AF 405 DBCO dye, I don't see a very clear distinction in groups. I am processing my samples using a Guava Easycyte flow cytometer and counter stain with SYBR Green I DNA Stain. I am wondering if anyone has prior experience with this dye and could give some insight.