Hoatlin:Crosslinking antibodies to beads

1. 400μL Protein A Sepharose Fast Flow Beads (GE Healthcare #17-1279-01)
2. Rotate @ room temp (RT) 30 min-3 hrs (3 hrs @ 4°C)
3. Wash 4X in PBS (1ml for each wach)
4. Wash 2x in 100mM Na Borate, pH 9.0
5. Resuspend beads in 1 ml 100mM Na Borate, pH 9.0 + 5.2mg DMP (Dimethyl pielimidate, Pierce #21667)
6. Rotate 45 min (don't exceed 1 hr) at RT
7. Spin 1 min at RT and recover beads
8. Wash beads 1x w/ 1M ethanolamine, pH 8.0 (Sigma #E9508)
9. Add 1 ml of 1M ethanolamine, pH 8.0
10. Rotate 2 hrs-overnight at 4°C
11. Wash 4x quickly in 100 mM glycin pH 3.0
12. Neutralize by washing 2x w/ PBS

• Store at 4°C
• Beads can be used several times