Hi all,
I'm going to do immunoprecipitation using lifetech Dynabeads Protein G.
I know for this experiment, I can incubate antibody and beads first then add to lysate or incubate antibody and lysate first then add beads.
My question is which method is better? The protocal come with the beads says incubate antibody and beads first. But in NOTES, it also says pre-incubate sample and antibody prior to bead capture can minimize nonspecific binding.
Thanks for your reply!
Hi there,
I don't think it actually makes any big difference. The only thing is that mixing beads with antibody first should mask some non specific sites on the beads. If you really want to lower non specific interaction you have to preclear your lysate with beads alone and/or irrelevant antibody alone first and then perform the experiment with the precleared lysate.
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