Using a mouse antibody for IP - should I use Dynabeads with Protein G or with sh - (May/11/2014 )
I plan to incorporate BrdU into the DNA of my proliferating human cells in culture, isolate the DNA and IP with an anti-BrdU antibody to capture those DNA sequences. I am wondering the disadvantages/advantages of using Dynabeads with either Protein G or sheep anti-mouse to capture the mouse anti-BrdU:DNA(BrdU) compexes? Specificity? Sensitivity? Efficiency? I feel that the two would be almost equal but I am not sure, which is why I am posting this question.
Most IPs are done with protein G - it gives you less IgG in the final solution and the interaction is with the "other" end of the antibody, so it doesn't interfere with the IP at all.