I am trying to measure the affinity of biotin to bind to streptavidin using AFM force spectroscopy. I would like to know the what buffer should be used and the pH to be maintained in which the reaction is happening inorder to measure the interaction.
The Streptavidin-biotin complex is generally stable over a wide pH ranges. However, the formation of the complex is disrupted only by conditions that lead to irreversible denaturation of the proteins.
Hi Alemu,
I am functionalizing the AFM tip with Streptavidin protein. I would like to check its resonant frequency in air after functionalization. I need to dry the tip for this purpose. Would the streptavidin still have the binding affinity for biotin when I place the tip back in the 1X PBS solution at pH 7.4?
Prefer using PBS buffer of pH 7.35 with approximately ionic strength of 0.5m NaCl, as the interaction is strong at this pH. As the pH becomes lower, the interaction decreases.
Hope this works for you.
Best wishes
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