Abstract
Since the discovery of the association of apolipoprotein E (apoE) 4 with Alzheimer’s disease 17 years ago, numerous in vitro experiments with the apoE isoforms (apoE2, apoE3, and apoE4) have been performed to try to understand the basis for this association. The majority of these studies used commercial sources for apoE, but some used recombinant protein. In either case, these studies were most often conducted without considering the ramifications of the structural and biophysical differences among the three isoforms or without adequate quality control of the preparations.
Here, we present a protocol for producing recombinant apoE that we have used successfully in our laboratory for the last 20 years. We also review the considerations that are critical for obtaining reliable and interpretable results with the end product.
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Acknowledgments
This work was supported in part by the National Institutes of Health/National Institute of Aging R01 AG028793; and The J. David Gladstone Institutes. The authors thank Gary Howard for editorial assistance, Linda Turney for manuscript preparation, and John Carroll for graphics assistance.
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Newhouse, Y., Weisgraber, K.H. (2010). Apolipoprotein E Expression and Purification. In: Roberson, E. (eds) Alzheimer's Disease and Frontotemporal Dementia. Methods in Molecular Biology, vol 670. Humana Press, Totowa, NJ. https://doi.org/10.1007/978-1-60761-744-0_10
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DOI: https://doi.org/10.1007/978-1-60761-744-0_10
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