How do I prepare immunoprecipitated (IP) samples for 2D DIGE?
If you use IP in your experiments, please do the IP and the washing the same way you did before. At the end, please transfer the beads to a 1.5 ml plastic tube, and wash the beads once with 0.5 ml of water (this should get rid of the salt which may interfere with IEF). Please remove water as completely as possible after the final wash. The bead pellets can be kept at -20C and sent to us on dry ice. We will elute protein complex from the beads using a buffer that contains 2 M thiourea, 7 M urea, 4% CHAPS and 30 mM This-HCl pH 8.8.
