1 reply to this topic

[ozgecanerdem]

Hi,

 

I want to prepare a lysis buffer for bacterial surface protein extraction. How can i prepare 8 M (Tris-buffered) urea containing 5 mM EDTA or is there anyone who have another idea for protein extraction?

[mdfenko]

you don't specify pH or buffer concentration or if any other components are required. so, here is the general procedure:

 

weigh out enough urea for the volume of medium you wish to prepare, add enough stock tris buffer and stock edta solution so that they will be at the necessary concentration when the volume is adjusted to the desired amount.

 

notes: the formula weight of urea is 60.06 gm/mole;

 

urea cools the solution as it dissolves, do not warm it! allow it to return to room temperature on its own. warming will cause the urea to decompose;

 

you will have to adjust to near final volume for the urea to fully solubilize.