Activated Immunobead® Matrix - Irvine Scientific

3. Incubate at 4°C for 1 hour.
4. Add 40.0 mg EDAC (1-ethyl-3 [3-dimethylaminopropyl] carbodiimide-
HCl) to the reaction mixture. Mix vigorously and completely.
5. Store at 4° C for at least 3 hours. The reaction mixture may be stored
overnight at this point, but if your protein is unstable, centrifuge and
wash at least once with PBS.
6. Proceed with the wash procedure (Section D).
C. Immunobead Preparation with Whole Antisera
Note: This procedure produces a lower specific activity than the
globulin procedure.
1. Centrifuge 200 µL of antisera at 5,000 x g to remove any insoluble
material.
2. Centrifuge the Immunobead suspension and discard the supernatant.
3. Add 100 µL antisera and 6.5 mL coupling buffer to Immunobead
pellet from Step 2. Re-suspend and mix well.
4. Incubate at 40° C for 1 hour.
5. Add 20.0 mg EDAC and mix vigorously. Maintain at 4° C for at least
3 hours.
6. Proceed with the wash sequence (Section D).
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