I attempted to dissolve 20 mM DTNB(5,5 - Dithiobis(2-nitrobenzoic acid) in 1X PBS buffer(0.1M pbs, pH 7.3), but it was unsuccessful.
Even after adding DTNB to the PBS buffer and vortexing for about 40 minutes, it did not dissolve. Then, I placed the DTNB-containing PBS buffer in a tumbler and reacted it for approximately 16 hours, but it still did not dissolve.
DTNB seems to be insoluble.....
Does anyone know a solution to this? I would appreciate any advice you may have.
If you are preparing 1 ml of total volume add 500 ul of ethanol to DTNB. It will dissolve completely by vortexing. Then add 1 % SDS (500 ul) to it. Vortex again. Prepare DTNB solution fresh every time.
Niti Sharma
thank you for your answer.
After dissolving DTNB, I plan to add purified protein to the solution. I'm concerned whether the ethanol and 1% SDS used to dissolve the DTNB might affect the protein
DTNB does not dissolve otherwise. You can optimize it for your protein by checking the activity.
DTNB is a compound that is slightly soluble in water. To ensure that DTNB can be fully dissolved, a buffer solution, such as a buffer with a pH value of approximately 8, is usually prepared first. Then, DTNB is mixed with sodium bicarbonate and dissolved using this buffer solution.
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