*Select ampholyte according to the range of IEF Chip to use. For ampholytes at 40% stock concentration, add 1 μL. For ampholytes at 100X, add 2 μL.
Dissolve the protein sample in the Working Rehydration Solution, prepared according to step 3. Sample may be diluted 2-fold or more with Working Rehydration Solution to reach desired protein concentration and decrease salt concentration.
A high salt concentration can affect protein separation during isoelectric focusing and cause the current to exceed 100 μA. Samples with a high salt concentration should be desalted by either:
NOTE: During protein resuspension, avoid heating proteins over 37 °C.
Quantify proteins. Between 0.1 - 100 μg of protein can be loaded for 2D electrophoresis. The optimum amount of protein will depend on detection method and sample complexity. The following protein amounts should be used as a starting point, users may need to optimize the loading amount for their particular sample.
Use the amount of protein loaded into Solution Chip to determine whether to use S, M, or L recipe type for protein separation.
The sample volume applied to the device is 13 - 15 μL. Sample can be diluted with Working Rehydration Solution, as necessary.
Select the appropriate power cord (provided with the device) that is compatible with the power outlets in your country. Firmly push the matching end into the back of the Auto2D® Device, and the other end into the power outlet.
The power switch is located next to the AC plug port on the back of the Auto2D® Device. Press the switch up to turn on.
The application will be automatically launched. From the screen, select Auto2D® Plus mode.
Load a Recipe
Choose SETTING > RECIPE. Touch the Recipe Information screen. Recipe select dialog box will appear.
Insert the Chips Into the Tray
On the Auto2D® Device touch screen, Choose OPEN > OK.
Once tray is open, place the PAGE Chip (light blue line) with the anode side in front. Place the Solution Chip Plus (dark blue lines) on top of the PAGE Chip with the cut-off corners in the front.
Apply the Solutions
*Wetted filter paper can be used as a wick to trap salt. Using filter paper is recommended to enhance separation of protein samples containing a high salt concentration when using Auto2D® Plus programs and is required when using Auto2D® Desalting program. Filter paper should be 0.2 – 0.3 mm in thickness and cut to a size of 8 mm x 4 mm.
Placement of the Electrode Chip Plus
Position the ▼ towards the front.
Remove the IEF Chip protective film.
Insert the IEF Chip in the slot indicated by the white dot on the Electrode Chip Plus.
Before the Starting Electrophoresis step, verify the following:
Choose CLOSE > OK > START > OK.
Electrophoresis will begin.
Remove the Gel
Choose OK > OPEN > OK.
Clean the Electrode Chip Plus with distilled water immediately and let it air dry after use.
Choose MENU > EXIT APPLICATION > OK > SYSTEM/OS SHUT DOWN > OK
Turn off switch on the back of the device.
Note: If condensation is observed inside the device, leave the tray open for a while before turning off the power, to allow the inside to dry.
Components exposed to samples should be disposed of with biological waste. Other materials should be disposed of according to all applicable international, federal, state, and local regulations. Refer to the components’ Safety Data Sheet for hazard information.