I think it should be fairly straightforward although the specifications of your assay might require specific modification of some steps such as blocking a bit. As usual, it's best to test the commercial and in-house side by side with the same material to see whether you are getting the same results. 

To begin, I would go for Nunc Maxisorb plates. Then, acquire purified streptavidin(you can ask for free samples from different suppliers to see what works and what does not), and test the saturation of the coating. I would start with 0.05 M Carbonate-Bicarbonate buffer (pH 9.6) and making a wide reciprocal dilution series of the SA to find the optimal or saturating coating concentration for your assay. You can also try coating on PBS as well, but higher pH usually works better for proteins.

For coating time and volume, I usually go for o/n at RT for antibodies/sugars/lipids at 100ul/well, or +4C for longer periods. Then wash with PBS/0.05% Tween20, block at RT for few hours with 150-200ul/well with PBS/0.5%BSA and you should be good to go with your assay.

Good luck!

I think it should be fairly straightforward although the specifications of your assay might require specific modification of some steps such as blocking a bit. As usual, it's best to test the commercial and in-house side by side with the same material to see whether you are getting the same results. 

To begin, I would go for Nunc Maxisorb plates. Then, acquire purified streptavidin(you can ask for free samples from different suppliers to see what works and what does not), and test the saturation of the coating. I would start with 0.05 M Carbonate-Bicarbonate buffer (pH 9.6) and making a wide reciprocal dilution series of the SA to find the optimal or saturating coating concentration for your assay. You can also try coating on PBS as well, but higher pH usually works better for proteins.

For coating time and volume, I usually go for o/n at RT for antibodies/sugars/lipids at 100ul/well, or +4C for longer periods. Then wash with PBS/0.05% Tween20, block at RT for few hours with 150-200ul/well with PBS/0.5%BSA and you should be good to go with your assay.

Good luck!

Hi Brian, make a 10ug/ml solution of streptavidin in H2O, put 100ul into the wells and leave then to dry ON. Wash and block with 0.1% BSA for 1 hr RT and you are ready to go.

Thanks Dr. Stefansson. I didn't realize it was that straightforward

Also, thank you Dr. Kotimaa. I didn't see your response at first.

can u apply the above method of immobilization of streptavidin to glass beads?

And do u know the rough estimation of the amount of streptavidin bound or the number of free biotin binding sites available per well

Cheers,

Yans