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I performed a test today with 100ug of protein. I was surprised that the beads are hardly visible. I am used to prot. G-sepharose beads and they are more visible than the GFP-Sepharose. Is that normal? As it is difficult to see them I am worried I don't get the IP protein or inespecific proteins are also there. Let me know when you have time if that is normal.
Asked on Apr 23 2012
The beads can appear to be translucent white when in the slurry. This is normal.Before taking out beads from the stock solution, it is important to mix the slurry well.
After the IP, when pelleting the bead-antibody-protein complex, the pellet should be clearly visible and stable so that the supernatant can be taken off without any bigger difficulties.
Please let me know how the results is looking on the WB. I am happy to help if optimisation is need.
Answered on Apr 23 2012