Advertisement

Detection of Cytokine Receptors Using Tyramide Signal Amplification for Immunofluorescence

  • Herui Wang
  • Ryan L. Pangilinan
  • Yan ZhuEmail author
Protocol
Part of the Methods in Molecular Biology book series (MIMB, volume 2108)

Abstract

Tyramide signal amplification (TSA) is an enzyme-mediated method to enhance the immunohistochemical detection of protein, nucleic acid, or other molecules in situ.

Here we describe immunofluorescent detection of a low-abundance cytokine receptor, interleukin-17 receptor B (IL17RB) in U2OS cells, using tyramide signal amplification. In addition, we present a tyramide signal amplification compatible double-color immunostaining protocol using primary antibodies from the same host species. Those applications allow detection of cellular proteins with enhanced sensitivity and add flexibility on primary antibody selection in double- or multicolor immunofluorescence staining assays.

Key words

Tyramide signal amplification Immunofluorescence Double-color immunostaining Interleukin-17 receptor B 

Notes

Acknowledgements

This work was supported by St. John’s University and NIH grant CA213426 to Yan Zhu.

References

  1. 1.
    Parra ER, Uraoka N, Jiang M, Cook P, Gibbons D, Forget MA, Bernatchez C, Haymaker C, Wistuba II, Rodriguez-Canales J (2017) Validation of multiplex immunofluorescence panels using multispectral microscopy for immune-profiling of formalin-fixed and paraffin-embedded human tumor tissues. Sci Rep 7(1):13380.  https://doi.org/10.1038/s41598-017-13942-8CrossRefPubMedPubMedCentralGoogle Scholar
  2. 2.
    Stack EC, Wang C, Roman KA, Hoyt CC (2014) Multiplexed immunohistochemistry, imaging, and quantitation: a review, with an assessment of Tyramide signal amplification, multispectral imaging and multiplex analysis. Methods 70(1):46–58.  https://doi.org/10.1016/j.ymeth.2014.08.016CrossRefPubMedPubMedCentralGoogle Scholar
  3. 3.
    Hunyady B, Krempels K, Harta G, Mezey E (1996) Immunohistochemical signal amplification by catalyzed reporter deposition and its application in double immunostaining. J Histochem Cytochem 44(12):1353–1362CrossRefGoogle Scholar
  4. 4.
    Monin L, Gaffen SL (2018) Interleukin 17 family cytokines: signaling mechanisms, biological activities, and therapeutic implications. Cold Spring Harb Perspect Biol 10(4):a028522.  https://doi.org/10.1101/cshperspect.a028522CrossRefPubMedPubMedCentralGoogle Scholar
  5. 5.
    Kolls JK, Linden A (2004) Interleukin-17 family members and inflammation. Immunity 21(4):467–476.  https://doi.org/10.1016/j.immuni.2004.08.018CrossRefPubMedGoogle Scholar
  6. 6.
    Alinejad V, Dolati S, Motallebnezhad M, Yousefi M (2017) The role of IL17B-IL17RB signaling pathway in breast cancer. Biomed Pharmacother 88:795–803.  https://doi.org/10.1016/j.biopha.2017.01.120CrossRefPubMedGoogle Scholar
  7. 7.
    Bie Q, Zhang B, Sun C, Ji X, Barnie PA, Qi C, Peng J, Zhang D, Zheng D, Su Z, Wang S, Xu H (2017) IL-17B activated mesenchymal stem cells enhance proliferation and migration of gastric cancer cells. Oncotarget 8(12):18914–18923.  https://doi.org/10.18632/oncotarget.14835CrossRefPubMedPubMedCentralGoogle Scholar
  8. 8.
    Wu HH, Hwang-Verslues WW, Lee WH, Huang CK, Wei PC, Chen CL, Shew JY, Lee EY, Jeng YM, Tien YW, Ma C, Lee WH (2015) Targeting IL-17B-IL-17RB signaling with an anti-IL-17RB antibody blocks pancreatic cancer metastasis by silencing multiple chemokines. J Exp Med 212(3):333–349.  https://doi.org/10.1084/jem.20141702CrossRefPubMedPubMedCentralGoogle Scholar
  9. 9.
    Ren L, Xu Y, Liu C, Wang S, Qin G (2017) IL-17RB enhances thyroid cancer cell invasion and metastasis via ERK1/2 pathway-mediated MMP-9 expression. Mol Immunol 90:126–135.  https://doi.org/10.1016/j.molimm.2017.06.034CrossRefPubMedGoogle Scholar

Copyright information

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

Authors and Affiliations

  1. 1.Department of Biological SciencesSt. John’s UniversityJamaicaUSA
  2. 2.Department of Biological SciencesSt. John’s UniversityQueensUSA

Personalised recommendations