Run the processor
1. Turn the power switch on the back of the unit to the "on" position. The display
will illuminate, and the moving parts will return to their home positions. After a
few seconds, the home screen will display.
2. Program or select your preferred western blot protocol (see “Program the
processor” on page 7).
3. Fill the buer bottle in the back of the processor. Ensure that the hose hanging
from the cap reaches the bottom of the bottle. Leave the cap loose to allow air
into the bottle as liquid is pumped out.
For each wash, 10–75 mL of wash buer can be dispensed for mini gels and
10–150 mL for midi gels. We recommend using 20–30 mL per wash for mini gels
and 30–50 mL for midi gels. Ensure there is enough wash buer to complete the
protocol.
4. Flush the buer lines by selecting Flush from the home screen menu and
then pressing Accept . Press Fill until wash buer is dispensed from
all nozzles. This may take several seconds. The wash buer will dispense into
the waste tray.
5. If using primary and secondary antibodies, 2 reagent tubes will be required
for each blot being processed. Check to ensure the foil on the bottom of the
reagent tubes is not damaged. Screw one reagent tube firmly and completely
into the primary antibody position, noted with the number 1 toward the back of
the slider, and the other into the secondary antibody position, noted with the
number 2, just behind the buer dispensing nozzle.
6. Pipet the appropriate antibody solution into each reagent tube.
A standard-sized mini blot processed in a mini tray requires a minimum of
3.5 mL of each primary antibody and secondary antibody solutions. A standard-
sized midi blot processed in a midi tray requires a minimum of 7.5 mL of each
primary antibody and secondary antibody solutions.
7. Place blot(s) into the sample tray protein side up with protein lanes
perpendicular to the tray gates. If using a mini tray, which has a central divider,
make sure the blot is on the same side of the divider as the appropriate antibody
reagent tubes.
8. Pour blocking buer of choice in the tray. Use no more than 45 mL of blocking
buer per mini tray and no more than 90 mL blocking buer for midi trays.
Replace the tray cover and make sure it is fully seated.
Tray Size Minimum Volume Maximum Volume
Mini 3.5 mL 45 mL
Midi 7.5 mL 90 mL
Bandmate™Automated Western Processor
Operate the Bandmate™Automated Western Blot Processor
Bandmate™Automated Western Blot Processor User Guide 9
