[Reagents and consumables needed for GST-fusion Protein Purification]

Provided in the kit:
Empty reagent cartridges
1.5 ml screw top tubes
Tips and tip sheathes
High salt solution
1 tube of 6X GST-Elution buffer
Cartridge template (GST-tag Purify & Pulldown)

[Preparation of bacterial lysate]

See page 2 of this manual.

[Preparation of 1X GST-Elution buffer]

1. Thaw 6X stock on ice.
2. Add 1.5 ml distilled water. This is sufficient for 6 samples.
3. Keep on ice before use. Freeze extra 1X Elution buffer at -80° C for later use for up to one week.

[Magnetic beads information]

1. For 1-4 ml protocol, use 40-80 µl of MagneGST^TM glutathione particles.
2. For 5-20 ml protocol, use 100-200 µl of MagneGST^TM glutathione particles.

[Preparation and filling of open cartridges]

1. Insert each reagent cartridge into
the 12GC cartridge rack until it
“clicks” into place.
2. Insert one or two cartridge templates labeled “GST-tag Purify & Pulldown” into empty slot(s) next to the open cartridges.

If 12 cartridges will be filled, place the cartridge template next to the rack.

3. Pipet 700 µl of Wash buffer into Well #2 through Well #4 of each cartridge (Fig. 1).

4. Pipet 1300 µl of Wash buffer into Well #5 of each cartridge (Fig. 1).

5. Pipet 350 µl of High salt solution into Well #6 of each cartridge (Fig. 1).

6. Once all cartridges are filled, remove the Cartridge template(s) from the 12GC cartridge rack and place the cartridge rack into the robot.

[Set up for the tip/tube rack]

1. Place tips, tubes, magnetic beads and sample in the tip/tube rack according to the following figure:

Row 4: 1.5 ml tube with sample: 0.6 ml for 1-4 ml protocol 1.2 ml for 5-20 ml protocol

Row 3: 1.5 ml tube with magnetic beads in 800 µl final volume diluted with distilled water.

Row 2: Tip in tip sheath
Row 1: 1.5 ml tube with 250 µl of 1X Elution buffer.

2. Make sure the caps have been removed from all tubes.

3. Place the tip/tube rack into the 12GC robot and close the door. The protocol is now ready to start.