Hello,

I am making cytoskeleton stabilizing buffer for the first time. The recipe I am using is 1 M NaCl, 100 mM PIPES, 30 mM MgCl2, 10 mM EGTA, 10 mM sucrose, in H2O. I made stock solutions of the MgCl2, EGTA, and sucrose. I diluted the MgCl2 and sucrose in water to the correct molarity and added solid NaCl. After the NaCl dissolved, I added solid PIPES. I made a mistake at this point--I got what I had read online mixed up and thought that PIPES needed an acidic pH to dissolve, so I added some HCl. It didn't seem to help it dissolve, so I went back to my references, realized my mistake, and then I added EGTA from my stock solution (I didn't add the EGTA in the beginning because I didn't want it to raise the pH of the solution and make it harder to dissolve the PIPES, so I figured I would add it in the end after dissolving PIPES, before adjusting the final pH and volume. Once I realized that I was mistaken about the pH PIPES needs, I added the EGTA). Then I started adding NaOH pellets. The PIPES quickly dissolved once I added the NaOH, but the whole solution turned bright yellow. The only clue I have found online so far is that NaOH produces a yellow color when it reacts with EtOH, but my solution does not contain EtOH so I don't think it could be that. Does anyone know what might have happened?

Thank you!