Standardized immunoprecipitation protocol for efficient isolation of native apolipoprotein E particles utilizing
Justin O'Leary1, Ana-Caroline Raulin1, Zonghua Li1
1Department of Neuroscience, Mayo Clinic, Jacksonville, FL 32224, USA.
None:
The apolipoprotein E protein (apoE) confers differential risk for Alzheimer's disease depending on which isoforms are expressed. Here, we present a 2-day immunoprecipitation protocol using the HJ15.4 monoclonal apoE antibody for the pull-down of native apoE particles. We describe major steps for apoE production via immortalized astrocyte culture and HJ15.4 antibody bead coupling for apoE particle pull-down, elution, and characterization. This protocol could be used to isolate native apoE particles from multiple model systems or human biospecimens.
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