His Monster Beads

PROTOCOL:

Binding:

1. Take 50-250 μL 20% His Monster beads Slurry (as provided) for 0.2-1 mg target protein purification,
equilibrated with 10 beads-volume binding buffer.
2. Remove liquid from the slurry, add ProteinFactory Rxn (or any other solution with target protein) con-
taining 0.2-1 mg target protein to the beads.
3. Re-suspend the beads and mix at 4-30 ℃ for 30 min.
4. Use magnet to suction the beads.
5. Discard the liquid.

Washing:

6. Add 1-1.2 mL washing buffer into beads, completely vortex for 3 to 5 min.
7. Magnetic suction the beads, remove all the liquid.
8. Repeat 6-7 for 3 times total.

Elution:

9. Add 5-10 beads-volume elution buffer , pipette 10 times.
10. Magnetic suction the beads and collect all liquid. Repeat 9 if needed.

Regeneration (if needed):

11. 50 beads-volume 2M NaOH resuspend the beads, vortex completely, rotate for 5 min,
or standing on table for 5min with occasionally vortex.
12. Magnetic suction the beads, remove all liquid.
13. Add 100 beads-volume H2O, vortex completely.
14.Magnetic suction the beads, remove all liquid.
15.Repeat 13,14 for 10 times total.

SOLUTION:

Binding buffer: 50 mM Tris-HCI pH 8.0, 500mM NaCI, 5 mM Imidazole;
Washing buffer: 50 mM Tris-HCI pH 8.0, 500mM NaCI, 20mM Imidazole;
Elution buffer: 50 mM Tris-HCI pH 8.0, 500mM NaCI, 250mM Imidazole;