SpringerLink
Log in

Application of Immunohistochemistry in Basic and Clinical Studies

  • Protocol
  • First Online:
Immune Mediators in Cancer

Part of the book series: Methods in Molecular Biology ((MIMB,volume 2108))

Abstract

Immunohistochemistry (IHC), also known as immunohistochemical staining, is an immune morphological analysis. It is a process of selectively identifying antigens (proteins) by antibodies in cells or tissue sections. This chapter introduces the procedure and application of immunohistochemistry. Although immunohistochemistry has a vast application in basic and clinical studies, this chapter focuses on its application in biomarker study, particularly in biomarkers that related to cancer diagnosis, prognosis, and drug development. Detail protocol of immunohistochemistry in formalin-fixed and paraffin-embedded tissue sections is included.

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

Access this chapter

Log in via an institution

Subscribe and save

Springer+ Basic
EUR 32.99 /Month
  • Get 10 units per month
  • Download Article/Chapter or Ebook
  • 1 Unit = 1 Article or 1 Chapter
  • Cancel anytime
Subscribe now

Buy Now

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 ship** worldwide - see info
Hardcover Book
USD 199.99
Price excludes VAT (USA)
  • Durable hardcover edition
  • Dispatched in 3 to 5 business days
  • Free ship** worldwide - see info

Tax calculation will be finalised at checkout

Purchases are for personal use only

Institutional subscriptions

Similar content being viewed by others

References

  1. Coons AH, Creech HJ, Jones RN (1941) Immunological properties of an antibody containing a fluorescent group. Proc Soc Exp Biol Med 47:200–202

    Article  CAS  Google Scholar 

  2. Childs GV (2014) History of immunohistochemistry. In: McManus LM, Mitchell RN (eds) Pathobiology of human disease. Elsevier, San Diego, pp 3775–3796

    Chapter  Google Scholar 

  3. Lindenmann J (1984) Origin of the terms “antibody” and “antigen”. Scand J Immunol 19(4):281–285

    Article  CAS  Google Scholar 

  4. The Nobel Prize in Physiology or Medicine (1901) nobelprize.org. https://www.nobelprize.org/prizes/medicine/1901/summary/

  5. The Nobel Prize in Physiology or Medicine (1908) nobelprize.org. https://www.nobelprize.org/prizes/medicine/1908/summary/

  6. Biomarkers Definitions Working Group (2001) Biomarkers and surrogate endpoints: preferred definitions and conceptual framework. Clin Pharmacol Ther 69(3):89–95

    Article  Google Scholar 

  7. Khleif SN, Doroshow JH, Hait WN, AACR-FDA-NCI Cancer Biomarkers Collaborative (2010) AACR-FDA-NCI. Cancer biomarkers collaborative consensus report: advancing the use of biomarkers in cancer drug development. Clin Cancer Res 16:3299–3318

    Article  CAS  Google Scholar 

  8. Smith NR, Womack C (2014) A matrix approach to guide IHC-based tissue biomarker development in oncology drug discovery. J Pathol 232(2):190–198

    Article  CAS  Google Scholar 

  9. Howat WJ, Lewis A, Jones P et al (2014) Antibody validation of immunohistochemistry for biomarker discovery: recommendations of a consortium of academic and pharmaceutical based histopathology researchers. Methods 70(1):34–38

    Article  CAS  Google Scholar 

  10. Shi SR, Liu C, Taylor CR (2007) Standardization of immunohistochemistry for formalin-fixed, paraffin-embedded tissue sections based on the antigen-retrieval technique: from experiments to hypothesis. J Histochem Cytochem 55(2):105–109

    Article  CAS  Google Scholar 

  11. Zu W, Li A, Chen T (2015) Rabbit monoclonal antibody. In: Fan L, Jeffrey P (eds) Handbook of practical immunohistochemistry: frequently asked questions, 2nd edn. Springer, New York

    Google Scholar 

  12. Schacht V, Kern JS (2015) Basics of immunohistochemistry. J Invest Dermatol 135:e30

    Article  Google Scholar 

  13. Werner M, Chott A, Fabiano A et al (2000) Effect of formalin tissue fixation and processing on immunohistochemistry. Am J Surg Pathol 24(7):1016–1019

    Article  CAS  Google Scholar 

  14. Grizzle WE (2009) Models of fixation and tissue processing. Biotech Histochem 84(5):185–193

    Article  CAS  Google Scholar 

  15. Taylor CR, Shi SR et al (1996) Comparative study of antigen retrieval heating methods: microwave, microwave and pressure cooker, autoclave, and steamer. Biotech Histochem 71(5):263–270

    Article  CAS  Google Scholar 

  16. Lott R, Tunnicliffe J, Sheppard E et al (2015) Practical guide to specimen handling in surgical pathology. College of American Pathologist, Northfield, IL

    Google Scholar 

  17. Yang DH, Smith ER, Roland IH, Sheng Z, He J, Martin WD, Hamilton TC, Lambeth JD, Xu XX (2002) Disabled-2 is essential for endodermal cell positioning and structural formation during mouse embryogenesis. Dev Biol 251:27–44

    Article  CAS  Google Scholar 

  18. Yang DH, Kasamo H, Miyauchi M, Tsuyama S, Murata F (1996) Ontogeny of sulfated glycoconjugate producing cells in the rat fundic gland. Histochem J 28:33–43

    Article  CAS  Google Scholar 

  19. Yang DH, Tsuyama S, Ge Y-B, Ohmori J, Wakamatsu D, Murata F (1997) Proliferation and migration kinetics of stem cells in the rat fundic gland. Histol Histopathol 12:717–728

    Google Scholar 

  20. Yang DH, Tsuyama S, Ohmori J, Hotta K, Katsuyama T, Murata F (1999) Phenotypic immunostaining of mucus-secreting cells of foregut origin. Acta Histochem Cytochem 32(2):135–140

    Article  Google Scholar 

  21. Yang DH, Tsuyama S, Murata F (2001) The expression of gastric H-K-ATPase mRNA and protein in develo** rat fundic gland. Histochem J 33:159–166

    Article  CAS  Google Scholar 

  22. Yang DH, Tsuyama S, Hotta K, Katsuyama T, Murata F (2000) Expression of N-acetylglucosamine residues in develo** rat fundic gland cells. Histochem J 32(3):187–193

    Article  CAS  Google Scholar 

  23. Yang DH, Moss EG (2003) Temporally regulated expression of Lin-28 in diverse tissue of the develo** mouse. Gene Expr Patterns 3(6):719–726

    Article  CAS  Google Scholar 

  24. Yang DH, Smith ER, Cohen C, Patriotis C, Godwin AK, Hamilton TC, Xu XX (2002) Molecular events associated with dysplastic morphological transformation and initiation of ovarian tumorigenicity. Cancer 94:2380–2392

    Article  CAS  Google Scholar 

  25. Yang DH, Fazili Z, Smith ER, Cai Q, Klein-Szanto A, Cohen C, Horowitz IR, Xu XX (2006) Disabled-2 heterozygous mice are predisposed to endometrial and ovarian tumorigenesis and exhibit sex-biased embryonic lethality in a p53 null background. Am J Pathol 169:258–267

    Article  CAS  Google Scholar 

  26. Yang DH, Cai KQ, Roland IH, Smith ER, Xu XX (2007) Disabled-2 is an epithelial surface positioning gene. J Biol Chem 282(17):13114–13122

    Article  CAS  Google Scholar 

  27. Yang DH, Smith ER, Cai KQ, Xu XX (2009) c-fos elimination compensates for disabled-2 requirement in mouse extraembryonic endoderm development. Dev Dyn 238(3):514–523

    Article  CAS  Google Scholar 

  28. Yang DH, McKee KK, Chen ZL, Mernaugh G, Strickland S, Zent R, Yurchenco PD (2011) Renal collecting system growth and function depend upon embryonic gamma1 laminin expression. Development 138(20):4535–4544

    Article  CAS  Google Scholar 

  29. Khan SA, Yang DH, Zhu F, Dubyk C, Tejani M, Cohen SJ, Hoffman JP, Burtness BA (2012) Survivin expression with resected pancreatic adenocarcinoma. Cancer Res 72(8):714–714

    Google Scholar 

  30. Keller LM, Galloway TJ, Holdbrook T, Ruth K, Yang DH, Dubyk C, Flieder D, Lango MN, Mehra R, Burtness B, Ridge JA (2013) p16 status, pathologic and clinical characteristics, biomolecular signature, and long term outcomes in unknown primary carcinomas of the head and neck. Head Neck 36(12):1677–1684

    Article  Google Scholar 

  31. Mekee KK, Yang DH, Patel R, Chen ZL, Strickland S, Takagi J, Sekiguchi K, Yurchenco PD (2012) Schwann cell myelination requires integration of laminin activities. J Cell Sci 125(Pt 19):4609–4619. https://doi.org/10.1242/jcs.107995. #equal contribution 1st author

    Article  CAS  Google Scholar 

  32. Sukhanova A, Gorin A, Serebriiskii IG, Gabitova L, Zheng H, Restifo D, Egleston BL, Cunningham D, Bagnyukova T, Liu H, Nikonova A, Adams GP, Zhou Y, Yang DH, Mehra R, Burtness B, Cai KQ, Klein-Szanto A, Kratz LE, Kelley RI, Weiner LM, Herman GE, Golemis EA, Astsaturov IA (2013) Targeting C4-demethylating genes in the cholesterol pathway sensitizes cancer cells to EGFR inhibitors via increased EGFR degradation. Cancer Discov 3(1):96–111. https://doi.org/10.1158/2159-8290.CD-12-0031.

    Article  CAS  PubMed  Google Scholar 

  33. Gabitova L, Restifo D, Gorin A, Manocha K, Handorf E, Yang DH, Cai K, Klein-Szanto AS, Cunningham D, Kratz L, Herman G, Golemis EA, Astsaturov I (2015) Endogenous sterol metabolites regulate growth of EGFR/KRAS-dependent tumors via LXR. Cell Rep 12(11):1927–1938. https://doi.org/10.1016/j.celrep.2015.08.023

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  34. Kudinov A, Deneka A, Nikonova A, Beck T, Ahn Y, Liu X, Yang DH, Golemis E, Boumber Y (2016) Musashi-2 (MSI2) supports TGF-β signaling and inhibits claudins to promote non-small cell lung cancer (NSCLC) metastasis. Proc Natl Acad Sci U S A 113(25):6955–6960. https://doi.org/10.1073/pnas.1513616113

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  35. Sodani K, Patel A, Anreddy N, Singh S, Yang DH, Kathawala RJ, Kumar P, Talele TT, Chen ZS (2014) Telatinib reverses chemotherapeutic multidrug resistance mediated by ABCG2 efflux transporter in vitro and in vivo. Biochem Pharmacol 89(1):52–61

    Article  CAS  Google Scholar 

  36. Mehra R, Zhu F, Yang DH, Cai KQ, Weaver J, Singh MK, Nikonova A, Golemis EA, Flieder DB, Cooper H, Lango MN, Ridge JA, Burtness B (2013) Quantification of excision repair cross complementing group 1 (ERCC1) and survival in p16-negative squamous cell head and neck cancers. Clin Cancer Res 19(23):6633–6643

    Article  CAS  Google Scholar 

  37. Egloff AM, Lee JW, Langer C, Quon H, Vaezi A, Grandis JR, Seethala RR, Wang L, Shin DM, Argiris A, Yang DH, Mehra R, Ridge JA, Patel UA, Burtness B, Forastiere AA (2014) Phase II study of cetuximab in combination with cisplatin and radiation in unresectable, locally advanced head and neck squamous cell carcinoma: Eastern Cooperative Oncology Group trial E3303. Clin Cancer Res 20(19):5041–5051

    Article  CAS  Google Scholar 

  38. Lee JW, Parameswaran J, Sandoval-Schaefer T, Eoh KJ, Yang DH, Zhu F, Mehra R, Sharma R, Gaffney SG, Perry EB, Townsend JP, Serebriiskii IG, Golemis EA, Issaeva N, Yarbrough WG, Koo JS, Burtness BA (2019) Combined Aurora kinase A (AURKA) and WEE1 inhibition demonstrates synergistic antitumor effect in squamous cell carcinoma of the head and neck. Clin Cancer Res 25(11):3430–3442

    Article  Google Scholar 

  39. Xu S, Yao H, Luo S, Zhang YK, Yang DH, Li D, Wang G, Hu M, Yao H∗, Wu X, Chen ZS, Xu J. (2017) A potent anticancer optimized from natural oridonin scaffold induces apoptosis and cell cycle arrest through the mitochondrial pathway, J Med Chem, 60(4):1449-1468. doi: https://doi.org/10.1021/acs.jmedchem.6b01652

    Article  CAS  PubMed  Google Scholar 

  40. Pileri SA, Roncador G et al (1997) Antigen retrieval techniques in immunohistochemistry: a comparison of different methods. J Pathol 183(1):116–123

    Article  CAS  Google Scholar 

  41. O’Leary TJ (2001) Standardization in immunohistochemistry. Appl Immunohistochem Mol Morphol 9(1):3–8

    PubMed  Google Scholar 

  42. Lin F, Chen Z (2014) Standardization of diagnostic immunohistochemistry: literature review and geisinger experience. Arch Pathol Lab Med 138(12):1564–1577

    Article  Google Scholar 

Download references

Author information

Authors and Affiliations

  1. Epitomics—An Abcam Company, Burlingame, CA, USA

    Aihua Li

  2. Department of Pharmacology, College of Pharmacy and Health Sciences, St. John’s University, Queens, NY, USA

    Dong-Hua Yang

Authors
  1. Aihua Li
    View author publications

    You can also search for this author in PubMed Google Scholar

  2. Dong-Hua Yang
    View author publications

    You can also search for this author in PubMed Google Scholar

Corresponding authors

Correspondence to Aihua Li or Dong-Hua Yang .

Editor information

Editors and Affiliations

  1. Department of Biological Sciences, St. John’s University, Queens, NY, USA

    Ivana Vancurova

  2. Department of Biological Sciences, St. John’s University, Queens, NY, USA

    Yan Zhu

Rights and permissions

Reprints and permissions

Copyright information

© 2020 Springer Science+Business Media, LLC, part of Springer Nature

About this protocol

Check for updates. Verify currency and authenticity via CrossMark

Cite this protocol

Li, A., Yang, DH. (2020). Application of Immunohistochemistry in Basic and Clinical Studies. In: Vancurova, I., Zhu, Y. (eds) Immune Mediators in Cancer. Methods in Molecular Biology, vol 2108. Humana, New York, NY. https://doi.org/10.1007/978-1-0716-0247-8_4

Download citation

  • DOI: https://doi.org/10.1007/978-1-0716-0247-8_4

  • Published:

  • Publisher Name: Humana, New York, NY

  • Print ISBN: 978-1-0716-0246-1

  • Online ISBN: 978-1-0716-0247-8

  • eBook Packages: Springer Protocols

Publish with us

Policies and ethics

Search

Navigation