Skip to main content
SpringerLink
Log in

Assaying Low-Density-Lipoprotein (LDL) Uptake into Cells

  • Protocol
  • First Online:
Cholesterol Homeostasis

Part of the book series: Methods in Molecular Biology ((MIMB,volume 1583))

Abstract

Determination of LDL particle uptake into cells is a valuable technique in the field of cholesterol metabolism. This allows assessment of LDL uptake capacity in different adherent and non-adherent cells types, as well as the effect of cellular, genetic, or pharmacological perturbations on this process. Here, we detail a general procedure that describes the production of fluorescently-labeled LDL particles and quantitative and non-quantitative assays for determining cellular LDL uptake.

This is a preview of subscription content, log in via an institution to check access.

Access this chapter

Log in via an institution
Protocol
USD 49.95
Price excludes VAT (USA)
  • Available as PDF
  • Read on any device
  • Instant download
  • Own it forever
eBook
USD 89.00
Price excludes VAT (USA)
  • Available as EPUB and PDF
  • Read on any device
  • Instant download
  • Own it forever
Softcover Book
USD 119.00
Price excludes VAT (USA)
  • Compact, lightweight edition
  • Dispatched in 3 to 5 business days
  • Free shipping worldwide - see info
Hardcover Book
USD 159.99
Price excludes VAT (USA)
  • Durable hardcover edition
  • Dispatched in 3 to 5 business days
  • Free shipping worldwide - see info

Tax calculation will be finalised at checkout

Purchases are for personal use only

Institutional subscriptions

References

  1. Maxfield FR, van Meer G (2010) Cholesterol, the central lipid of mammalian cells. Curr Opin Cell Biol 22:422–429

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  2. Hegele RA (2009) Plasma lipoproteins: genetic influences and clinical implications. Nat Rev Genet 10:109–121

    Article  CAS  PubMed  Google Scholar 

  3. Mozaffarian D et al (2015) Heart disease and stroke statistics--2015 update: a report from the American Heart Association. Circulation 131:e29–322

    Article  PubMed  Google Scholar 

  4. Brown MS, Goldstein JL (1986) A receptor-mediated pathway for cholesterol homeostasis. Science 232:34–47

    Article  CAS  PubMed  Google Scholar 

  5. Hobbs HH, Russell DW, Brown MS, Goldstein JL (1990) The LDL receptor locus in familial hypercholesterolemia: mutational analysis of a membrane protein. Annu Rev Genet 24:133–170

    Article  CAS  PubMed  Google Scholar 

  6. Goldstein JL, DeBose-Boyd RA, Brown MS (2006) Protein sensors for membrane sterols. Cell 124:35–46

    Article  CAS  PubMed  Google Scholar 

  7. Zelcer N, Hong C, Boyadjian R, Tontonoz P (2009) LXR regulates cholesterol uptake through Idol-dependent ubiquitination of the LDL receptor. Science 325:100–104

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  8. Sorrentino V, Zelcer N (2012) Post-transcriptional regulation of lipoprotein receptors by the E3-ubiquitin ligase inducible degrader of the low-density lipoprotein receptor. Curr Opin Lipidol 23:213–219

    Article  CAS  PubMed  Google Scholar 

  9. Abifadel M et al (2003) Mutations in PCSK9 cause autosomal dominant hypercholesterolemia. Nat Genet 34:154–156

    Article  CAS  PubMed  Google Scholar 

  10. Cohen J et al (2005) Low LDL cholesterol in individuals of African descent resulting from frequent nonsense mutations in PCSK9. Nat Genet 37:161–165

    Article  CAS  PubMed  Google Scholar 

  11. Blom DJ et al (2014) A 52-week placebo-controlled trial of evolocumab in hyperlipidemia. N Engl J Med 370:1809–1819

    Article  CAS  PubMed  Google Scholar 

  12. Stein EA et al (2012) Effect of a monoclonal antibody to PCSK9 on LDL cholesterol. N Engl J Med 366:1108–1118

    Article  CAS  PubMed  Google Scholar 

  13. Stein EA, Raal F (2014) Reduction of low-density lipoprotein cholesterol by monoclonal antibody inhibition of PCSK9. Annu Rev Med 65:417–431

    Article  CAS  PubMed  Google Scholar 

  14. Kleinveld HA, Duif PF, Pekelharing HL, van Rijn HJ (1996) Oxidation of lipoprotein(a) and low density lipoprotein containing density gradient ultracentrifugation fractions. Biochim Biophys Acta 1303:15–21

    Article  PubMed  Google Scholar 

  15. Levels JHM, Lemaire LCJM, van den Ende AE, van Deventer SJH, van Lanschot JJB (2003) Lipid composition and lipopolysaccharide binding capacity of lipoproteins in plasma and lymph of patients with systemic inflammatory response syndrome and multiple organ failure. Crit Care Med 31:1647–1653

    Article  CAS  PubMed  Google Scholar 

  16. Motazacker MM et al (2012) Advances in genetics show the need for extending screening strategies for autosomal dominant hypercholesterolaemia. Eur Heart J 33:1360–1366

    Article  CAS  PubMed  Google Scholar 

  17. Redgrave TG, Roberts DC, West CE (1975) Separation of plasma lipoproteins by density-gradient ultracentrifugation. Anal Biochem 65:42–49

    Article  CAS  PubMed  Google Scholar 

Download references

Acknowledgment

We thank Dr. Geesje M. Dallinga-Thie (AMC, The Netherlands) for her advice and comments. AL is supported by a Dekker grant from the Dutch Heart Foundation (2016T015). NZ is an Established Investigator of the Dutch Heart Foundation (2013T111) and is supported by an ERC Consolidator grant (617376) from the European Research Council.

Author information

Authors and Affiliations

  1. Department of Medical Biochemistry, Academic Medical Center, University of Amsterdam, Meibergdreef 15, Amsterdam, 1105AZ, The Netherlands

    Anke Loregger, Jessica K. Nelson & Noam Zelcer

Authors
  1. Anke Loregger
    View author publications

    You can also search for this author in PubMed Google Scholar

  2. Jessica K. Nelson
    View author publications

    You can also search for this author in PubMed Google Scholar

  3. Noam Zelcer
    View author publications

    You can also search for this author in PubMed Google Scholar

Corresponding authors

Correspondence to Anke Loregger or Noam Zelcer .

Editor information

Editors and Affiliations

  1. Faculty of Pharmacy, The University of Sydney, Sydney, New South Wales, Australia

    Ingrid C. Gelissen

  2. School of Biotechnology and Biomolecular Sciences, The University of New South Wales, Sydney, New South Wales, Australia

    Andrew J. Brown

Rights and permissions

Reprints and permissions

Copyright information

© 2017 Springer Science+Business Media LLC

About this protocol

Cite this protocol

Loregger, A., Nelson, J.K., Zelcer, N. (2017). Assaying Low-Density-Lipoprotein (LDL) Uptake into Cells. In: Gelissen, I., Brown, A. (eds) Cholesterol Homeostasis. Methods in Molecular Biology, vol 1583. Humana Press, New York, NY. https://doi.org/10.1007/978-1-4939-6875-6_5

Download citation

  • DOI: https://doi.org/10.1007/978-1-4939-6875-6_5

  • Published:

  • Publisher Name: Humana Press, New York, NY

  • Print ISBN: 978-1-4939-6873-2

  • Online ISBN: 978-1-4939-6875-6

  • eBook Packages: Springer Protocols

Publish with us

Policies and ethics

Search

Navigation