Activated Immunobead® Matrix - Irvine Scientific

1. Divide the material into equal portions in centrifuge tubes. A wash
volume of at least 80 mL/200 mg Immunobead reagent is required.
2. Fill tubes to capacity and balance the tubes with PBS. Centrifuge
at a minimum of 1,000 x g for at least 10 minutes.
3. Immediately after the centrifuge stops, decant and discard the
supernatant by completely inverting the tubes.
4. Add a small volume of PBS to each tube. Re-suspend the pellet by
using a vortex mixer or vigorous mixing.
5. Repeat steps 2 and 3.
6. Re-suspend the pellet in a small volume of 1.4 M NaCl-PBS. Fill the
tube(s) to capacity and balance with 1.4 M NaCl-PBS.
7. Centrifuge at 1,000 x g for 10 minutes.
8. Immediately after the centrifuge stops, decant the supernatant with
a single complete inversion of the tubes. Drain briefly.
9. Repeat steps 6, 7, and 8.
10. Wash twice with PBS. (Repeat twice Steps 4, 2, and 3.)
11. Re-suspend in PBS and allow to stand on ice (4°C) for at least 3
hours to allow complete renaturation of bound antibody.
12 a.) At this point, the reagent may be used. Wash twice as above
and combine to a final volume of 20 mL (10 mg/mL of Immunobead
reagent).
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