During acetylcholinesterase assay I used a inhibitor for finding its inhibitory activity, during dissolving the compound in buffer the solution becomes like there is some dust in there with it. I tried to centrifuge but it does not become settled. Can I take OD from this solution which is added with acetylcholinesterase to find it inhibitory effect. Will there be any effect due to these dust like particles?
As for the influence of dust like particles on inhibition of acetylcholinesterases no positive or negative answer can be expected. An experiment only can give you the correct answer.
Since you do not characterize neither your inhibitor /properties/ nor the buffer, I guess you added it to buffer in the form of powder.
If it is possible, dissolve the inhibitor in your buffer solution, and try to filter it under a vacuum through a very dense filter. Maybe it will work. Centrifuge cannot separate your dust like particles from the buffer in the case of identical or almost identical densities of both inhibitor and buffer solution.
Does not your inhibitor interact with any component of the buffer solution thus forming a precipitate? When it does not, try another approache:
In the case of limited solubility or slow dissolution of an inhibitor, try to dissolve smaller amount of it in distilled water, and look for the dust like particles formation. If there are not observable any, add the inhibitor to the buffer in the form of water solution. This might work.
Best regards,
R.S.
It may be that you have exceeded the solubility of your compound in the buffer, and the dust-like particles are the insoluble inhibitor. If this is the case, you may be able to improve the solubility of the inhibitor by changing th pH or reducing the salt concentration.
The OD will be elevated by scattering from the particles.
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