How much Tris do I need to get pH neutral glycine - (Jan/18/2008 )

I have eluted my protein with 1ml of pH 2 glycine and I want to bring it back to pH neutral before denaturing and running on a gel. I have Tris at pH 8.8, so how do I go about calculating how much of that I need to add to bring it to pH neutral. (I could use highe pH tris as well if that helps - ideally I want to add a very small volume).

---

you can add 100µL of 1M Tris pH 7.5.

---

Surely 1ml of pH2 50mM glycine plus 100ul of pH 7.5 1M tris would still be acidic?

If not could you let me know how you worked it out?

Thanks

---

the procedure i used was to add 1/20th fraction volume of 1M tris pH 8 (for glycine pH 2.5) to the tube, then drip the fraction into the buffer "cushion".

---

Can I ask if anyone knows how to do the calculation to know that this will give a final solution that is pH 7.4?

Thanks, any help will be greatly appreciated

---

all I know is that I use Glycine pH2,5 and add 100µL of Tris 1M pH 7.5 or 8 to 1ml aliquot. I never tested the final pH, you can try it out though before eluting your protein.

I don't know how to do the calculation.

---

instead of calculating, why not actually determine it.

take the buffer that you will use to elute the column and fill a tube with as much as you will find in your fractions. then add 1M tris until you reach your preferred pH. put that amount into each tube.

---