Skip to main content

Global Detection of Proteins by Label-Based Antibody Array

  • Protocol
  • First Online:
Antibody Arrays

Part of the book series: Methods in Molecular Biology ((MIMB,volume 2237))

  • 1075 Accesses

Abstract

Because of narrow availability of antibody pairs and potential cross-reactivity between antibodies, the development of sandwich-based antibody arrays which need a pair of antibodies for each target has been restricted to higher density resulting in limited proteomic breadth of detection. Label-based array is one way to overcome this obstacle by directly labeling all targets in samples with fluorescent dyes such as Cy3 and Cy5. The labeled samples are then applied on the antibody array chip composed of capture antibodies. In this chapter, we will introduce this technology including array production and sample detection assay.

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

Access this chapter

Protocol
USD 49.95
Price excludes VAT (USA)
  • Available as PDF
  • Read on any device
  • Instant download
  • Own it forever
eBook
USD 99.00
Price excludes VAT (USA)
  • Available as EPUB and PDF
  • Read on any device
  • Instant download
  • Own it forever
Softcover Book
USD 129.99
Price excludes VAT (USA)
  • Compact, lightweight edition
  • Dispatched in 3 to 5 business days
  • Free shipping worldwide - see info
Hardcover Book
USD 199.99
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. Hashimoto I, Wada J, Hida A, Baba M, Miyatake N, Eguchi J, Shikata K, Makino H (2006) Elevated serum monocyte chemoattractant protein-4 and chronic inflammation in overweight subjects. Obesity (Silver Spring) 14(5):799–811

    Article  CAS  Google Scholar 

  2. Ray S, Britschgi M, Herbert C, Takeda-Uchimura Y, Boxer A, Blennow K, Friedman LF, Galasko DR, Jutel M, Karydas A, Kaye JA, Leszek J, Miller BL, Minthon L, Quinn JF, Rabinovici GD, Robinson WH, Sabbagh MN, So YT, Sparks DL, Tabaton M, Tinklenberg J, Yesavage JA, Tibshirani R, Wyss-Coray T (2007) Classification and prediction of clinical Alzheimer’s diagnosis based on plasma signaling proteins. Nat Med 13(11):1359–1362

    Article  CAS  Google Scholar 

  3. Turtinen LW, Prall DN, Bremer LA, Nauss RE, Hartsel SC (2004) Antibody array-generated profiles of cytokine release from THP-1 leukemic monocytes exposed to different amphotericin B formulations. Antimicrob Agents Chemother 48(2):396–403

    Article  CAS  Google Scholar 

  4. De CF, Dassencourt L, Anract P (2004) The inflammatory side of human chondrocytes unveiled by antibody microarrays. Biochem Biophys Res Commun 323(3):960–969

    Article  Google Scholar 

  5. Haddad G, Belosevic M (2009) Transferrin-derived synthetic peptide induces highly conserved pro-inflammatory responses of macrophages. Mol Immunol 46(4):576–586

    Article  CAS  Google Scholar 

  6. Argas DL, Nascimbene C, Krishnan C, Zimmerman AW, Pardo CA (2005) Neuroglial activation and neuroinflammation in the brain of patients with autism. Ann Neurol 57(1):67–81

    Article  Google Scholar 

  7. Liu BC, Zhang L, Lv LL, Wang YL, Liu DG, Zhang XL (2006) Application of antibody array technology in the analysis of urinary cytokine profiles in patients with chronic kidney disease. Am J Nephrol 26(5):483–490

    Article  CAS  Google Scholar 

  8. Fu SY, Su GW, McKinley SH, Yen MT (2007) Cytokine expression in pediatric subperiosteal orbital abscesses. Can J Ophthalmol 42(6):865–869

    Article  Google Scholar 

  9. Coppinger JA, O’Connor R, Wynne K, Flanagan M, Sullivan M, Maguire PB, Fitzgerald DJ, Cagney G (2007) Moderation of the platelet releasate response by aspirin. Blood 109(11):4786–4792

    Article  CAS  Google Scholar 

  10. Simcock DE, Kanabar V, Clarke GW, O’Connor BJ, Lee TH, Hirst SJ (2007) Proangiogenic activity in bronchoalveolar lavage fluid from patients with asthma. Am J Respir Crit Care Med 176(2):146–153

    Article  CAS  Google Scholar 

  11. Kim HB, Kim CK, Iijima K, Kobayashi T, Kita H (2009) Protein microarray analysis in patients with asthma: elevation of the chemokine PARC/CCL18 in sputum. Chest 135(2):295–302

    Article  CAS  Google Scholar 

  12. Matsunaga K, Yanagisawa S, Ichikawa T, Ueshima K, Akamatsu K, Hirano T, Nakanishi M, Yamagata T, Minakata Y, Ichinose M (2006) Airway cytokine expression measured by means of protein array in exhaled breath condensate: correlation with physiologic properties in asthmatic patients. J Allergy Clin Immunol 118(1):84–90

    Article  CAS  Google Scholar 

  13. Sack RA, Conradi L, Krumholz D, Beaton A, Sathe S, Morris C (2005) Membrane array characterization of 80 chemokines, cytokines, and growth factors in open- and closed-eye tears: angiogenin and other defense system constituents. Invest Ophthalmol Vis Sci 46(4):1228–1238

    Article  Google Scholar 

  14. Kverka M, Burianova J, Lodinova-Zadnikova R, Kocourkova I, Cinova J, Tuckova L, Tlaskalova-Hogenova H (2007) Cytokine profiling in human colostrum and milk by protein array. Clin Chem 53(5):955–962

    Article  CAS  Google Scholar 

  15. Fujita K, Ewing CM, Sokoll LJ, Elliott DJ, Cunningham M, De Marzo AM, Isaacs WB, Pavlovich CP (2008) Cytokine profiling of prostatic fluid from cancerous prostate glands identifies cytokines associated with extent of tumor and inflammation. Prostate 68(8):872–882

    Article  CAS  Google Scholar 

  16. Lin Y, Huang R, Chen LP, Lisoukov H, Lu ZH, Li S, Wang CC, Huang RP (2003) Profiling of cytokine expression by biotin label-based protein arrays. Proteomics 3(9):1750–1757

    Article  CAS  Google Scholar 

Download references

Author information

Authors and Affiliations

Authors

Corresponding author

Correspondence to Zhizhou Kuang .

Editor information

Editors and Affiliations

Rights and permissions

Reprints and permissions

Copyright information

© 2021 Springer Science+Business Media, LLC, part of Springer Nature

About this protocol

Check for updates. Verify currency and authenticity via CrossMark

Cite this protocol

Kuang, Z., Chen, LP., Huang, R., Huang, RP. (2021). Global Detection of Proteins by Label-Based Antibody Array. In: Whittaker, K.C., Huang, RP. (eds) Antibody Arrays. Methods in Molecular Biology, vol 2237. Humana, New York, NY. https://doi.org/10.1007/978-1-0716-1064-0_4

Download citation

  • DOI: https://doi.org/10.1007/978-1-0716-1064-0_4

  • Published:

  • Publisher Name: Humana, New York, NY

  • Print ISBN: 978-1-0716-1063-3

  • Online ISBN: 978-1-0716-1064-0

  • eBook Packages: Springer Protocols

Publish with us

Policies and ethics