How much dialysis solution? - (Apr/13/2006 )
Hello,
I am purifying an ADH enzyme from a bacterial cell culture. I have just finished my ammonium sulfate washes and am getting ready to go into dialysis. I found a formula for the amount of buffer to use, but i need to figure out the concentration of the salt in the enzyme solution.
The steps for the ammonium sulfate are: bring to 35% (6.5184g), spin and remove precipitate, bring to 65% (additional 6.634g) and keep precipitate. Dissolve precipitate in 20mM tris hcl buffer, ph7.2 and dialyse.
any suggestions on amount of tris i should dissolve the precipitate in? and possible salt concentration? Thank you
Cheers
dez
-dez-
QUOTE (dez @ Apr 13 2006, 07:04 PM)
Hello,
I am purifying an ADH enzyme from a bacterial cell culture. I have just finished my ammonium sulfate washes and am getting ready to go into dialysis. I found a formula for the amount of buffer to use, but i need to figure out the concentration of the salt in the enzyme solution.
The steps for the ammonium sulfate are: bring to 35% (6.5184g), spin and remove precipitate, bring to 65% (additional 6.634g) and keep precipitate. Dissolve precipitate in 20mM tris hcl buffer, ph7.2 and dialyse.
any suggestions on amount of tris i should dissolve the precipitate in? and possible salt concentration? Thank you
Cheers
dez
I am purifying an ADH enzyme from a bacterial cell culture. I have just finished my ammonium sulfate washes and am getting ready to go into dialysis. I found a formula for the amount of buffer to use, but i need to figure out the concentration of the salt in the enzyme solution.
The steps for the ammonium sulfate are: bring to 35% (6.5184g), spin and remove precipitate, bring to 65% (additional 6.634g) and keep precipitate. Dissolve precipitate in 20mM tris hcl buffer, ph7.2 and dialyse.
any suggestions on amount of tris i should dissolve the precipitate in? and possible salt concentration? Thank you
Cheers
dez
I would use as few as possible, not to dilute too much your protein. It depends on the size of your precipitate.
-Missele-