I have biotin-labeled substrates and I want to visualize them under the microscope. I'm thinking of immobilizing streptavidin on slides or coverslips and then incubate them with my substrates. I'm looking for glass slides with streptavidin on them and all I find is microarray supplies and they are quite expensive. Any protocol/suggestions on how to immobilize streptavidin on slides?
u can coat your slides or tissue culture wells with need concentration of streptavidin in 0.1M carbonate buffer (NaHCO3), coating should be done overnight at 4 degree and subsequent washes to remove excess uncoated fraction.
Before placing streptavidin on the slide (the way Ankit suggested) I advise to pre-coat the glass slide with APTES. This ensures a good chemical bond between the glass and your molecules. Personally, I was able to bind, BSA, HSA, biotin and steptavidin,etc on these glasses and I was sure it stayed there.
Do you add a coverslip on top of the streptavidin coated slide or keep it in a moist environment or do you just let it dry completely during overnight incubation to reach a good coating?
Is there a more complete protocol for this? (i.e. how many wash steps/in what, etc.?) I'd really like to do this, but I'm very new to coating materials on glass.
I am really interested in getting a detailed protocol as well. Anyone has it? Thanks
Benjamin Samuel Goldschmidt?
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