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X-tal protocols

Equilibrium Dialysis with microdialysis cells.

Protein Detergent Methods Euipment Screen Drops Reference
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Micro-diffusion Cells.

dialysis button
M
P
O
D
T
- Microdialysis button
- Protein
- Outer solution
- Dialysis membrane
- Tubing

Buttons made out of glass, perspex or Teflon.

The volume of the cell is between 5 and 350 microliter.

(A) The cell is filled with the protein sample.

(B) The applicator with O-ring and the dialysis membrane is placed quickly over the cell, to prevend air bubbles in the cell.

(C) The dialysis membrane is fixed in place with the O-ring.

(D) The applicator is removed and the button in placed in a well filled with mother liquor. The well is closed with a coverslide or sealing tape.


Using the dialysis button

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Double dialysis method.

Double dialysis with button
M
P
O
I
D
T
- Microdialysis button
- Protein
- Outer solution
- Intermediate solution
- Dialysis membrane
- Tubing

A button containing the protein solution, sealed with a dialysis membrane, is placed in a small vail containing an intermediate solution, with a precipitant concentration insufficient to crystallize the protein. This vial is sealed with a dialysis membrane and placed in a larger vial which contains the outer solution, containing precipitant at a higher concentration which is sufficient to precipitate the protein. The rate of dialysis is varied by changing the relative volumes of the compartments and the solution concentrations.[Thomas, 1989]


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