# some protocol - buffer solution (Aug/23/2006 )

I need to prepare the solution (50 mM Tris-HCl, pH 8.0, 500 mM KCl, 2 mM DTT, 1 mM EDTA, 1 mM orthovanadate, and 10% glycerol) Do you have some protocol or sell the prepared solution?

-zacosa-

QUOTE (zacosa @ Aug 23 2006, 02:12 PM)
(50 mM Tris-HCl, pH 8.0, 500 mM KCl, 2 mM DTT, 1 mM EDTA, 1 mM orthovanadate, and 10% glycerol)

This IS the protocol. Or am I missing something?

-Jou-

what's your problem? to prepare tris-HCL?
I would prepare either a 1M or 0.1M tris-HCL.
Let's do a 0.1M.
the molecular weight of tris is 121.1
so to prepare 1L of a 0.1M you have to weight 12.11g, solubilize it in 800 mL, with the help of a pH-meter, add enough HCL concentrated to reach a pH of 8.0.

or is it to calculate the amount of powder to make a x molar solution?

you need to know the molecular weight (it's written on the bottle)
it means that 1 mol weight x grams.

so, to do 1L of 1M solution you have to weight x grams.

for the glycerol, is it 10% v/v of wt/v ?
usually it's v/v, so you have to take 10 mL of glycerol for a 100 mL solution
if it's wt/v, it means 10 grams per 100 mL .
If I remember well the density is 1.25g/mL, so you use 8 mL of glycerol per 100 mL solution

hope it helps

-Missele-