My substrate is biotinylated and I need to immobilize it on streptavidin slides. However, I'm not sure what conc. or how much to use and how long to incubate. By the way, it needs to be kept cold, so room temperature conditions are not an option. And also, what about the wash and blocking conditions? How long should they be? I can not use any detergent in wash or blocking buffers and I'm planning on using PBS and 1% BSA.
This depends on a ton of variables, so can you give more details about your system. How are you measuring binding, if at all? What type of substrate (just glass, yes?).
In terms of blocking, it depends on the application. BSA may work, but you also can try cassein or dehydrated milk, or even a commercial blocker like SuperBlock.
Many different conditions might be suitable. However, avoid natural blocking agents, as long as they are not definitely specified as "biotin-free". Any biotin traces in your reagents or buffers will "inactivate" your streptavidin irreversibly.
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