39
1. Place perpendicularly Teflon bar with two connecting rods at
each edge of the bar.
2. Place precut soaked membrane on the Teflon bar. Ensure that
membrane below the top edge of the bar and the lower edge of
membrane covers the bottom of all wells.
3. Repeat layering soaked membrane and Teflon bar until the
device is fully assembled (see Note 8 and Fig. 2). Flatten membrane before the next Teflon bar is put in place.
4. Insert the fully assembled Teflon block into the base of dialysis
device and then tighten them.
5. Immediately add receptor solution to the dialysis side of the
wells, assigned as receptor compartment, to prevent dehydration of soaked membranes. Ensure that no bubbles or leakage is
noticed.
6. Add equal volume of test solutions to the other side of the dialysis well (as donor compartment) using appropriate pipetting
device (see Note 9) and start timer.
7. Cover the top surface of dialysis device with an adhesive sealing
film to prevent evaporation.
8. At equilibrium time (see Note 10), discard solutions from
receptor compartment to analyze the amount of aggregating
molecule with an appropriate analytical method.
1. Add 1.2 mL of receptor medium to 1.5 mL conical tube and set
aside.
2. Load ultrapure water into dialysis device and observe for at least
5 min to moisture membrane and check the integrity of device.
If droplets are noticed from across the membrane, leakage
occurs and the device should not be used.
3. Decant ultrapure water and shake dialysis device to completely
remove water. Do not touch the wetted membrane with
ungloved hands. Once the membrane is wet, do not let it
become dry.
4. Immediately pipette a 0.5 mL of tested solution and then cap
the dialysis device to prevent evaporation.
5. Slowly place the filled dialysis device into conical tube containing receptor (i.e., dialysis cell). Ensure that the membrane contacts with receptor solution and does not introduce any
bubbles.
6. Place individually dialysis cell on microcentrifuge tube rack or
suitable supportive apparatus.
7. Shake gently on an orbital shaker (i.e., 100 rpm). Experiment is
performed at room temperature (unless indicated otherwise).
3.1.2 Micro-Equilibrium
Dialysis Device
3.1.3 Cuplike MINI
Dialysis Device
Aggregate Determination by Permeation Technique
1. Place perpendicularly Teflon bar with two connecting rods at
each edge of the bar.
2. Place precut soaked membrane on the Teflon bar. Ensure that
membrane below the top edge of the bar and the lower edge of
membrane covers the bottom of all wells.
3. Repeat layering soaked membrane and Teflon bar until the
device is fully assembled (see Note 8 and Fig. 2). Flatten membrane before the next Teflon bar is put in place.
4. Insert the fully assembled Teflon block into the base of dialysis
device and then tighten them.
5. Immediately add receptor solution to the dialysis side of the
wells, assigned as receptor compartment, to prevent dehydration of soaked membranes. Ensure that no bubbles or leakage is
noticed.
6. Add equal volume of test solutions to the other side of the dialysis well (as donor compartment) using appropriate pipetting
device (see Note 9) and start timer.
7. Cover the top surface of dialysis device with an adhesive sealing
film to prevent evaporation.
8. At equilibrium time (see Note 10), discard solutions from
receptor compartment to analyze the amount of aggregating
molecule with an appropriate analytical method.
1. Add 1.2 mL of receptor medium to 1.5 mL conical tube and set
aside.
2. Load ultrapure water into dialysis device and observe for at least
5 min to moisture membrane and check the integrity of device.
If droplets are noticed from across the membrane, leakage
occurs and the device should not be used.
3. Decant ultrapure water and shake dialysis device to completely
remove water. Do not touch the wetted membrane with
ungloved hands. Once the membrane is wet, do not let it
become dry.
4. Immediately pipette a 0.5 mL of tested solution and then cap
the dialysis device to prevent evaporation.
5. Slowly place the filled dialysis device into conical tube containing receptor (i.e., dialysis cell). Ensure that the membrane contacts with receptor solution and does not introduce any
bubbles.
6. Place individually dialysis cell on microcentrifuge tube rack or
suitable supportive apparatus.
7. Shake gently on an orbital shaker (i.e., 100 rpm). Experiment is
performed at room temperature (unless indicated otherwise).
3.1.2 Micro-Equilibrium
Dialysis Device
3.1.3 Cuplike MINI
Dialysis Device
Aggregate Determination by Permeation Technique