Abstract
The in situ expression levels and subcellular localization of molecules involved in signal transduction using specific antibodies can be useful for prognosis and diagnosis of human diseases such as cancer. In addition, it has the potential to be helpful in monitoring biologic response to targeted therapies. The increasing availability of such antibodies makes these studies feasible. However, compared to typical immunohistochemical stains in which stabile molecules such as cytokeratins are targeted, additional validation may be required for signal transduction immunohistochemistry.
Access this chapter
Tax calculation will be finalised at checkout
Purchases are for personal use only
Similar content being viewed by others
References
Mandell JW. (2003) Phosphorylation state-specific antibodies: applications in investigative and diagnostic pathology. Am J Pathol 163, 1687–1698.
Ross AH, Baltimore D, Eisen HN. (1981) Phosphotyrosine-containing proteins isolated by affinity chromatography with antibodies to a synthetic hapten. Nature 294, 654–656.
Glenney JR, Jr., Zokas L, Kamps MP. (1988) Monoclonal antibodies to phosphotyrosine.J Immunol Methods 109, 277–285.
Heffetz D, Fridkin M, Zick Y. (1991) Generation and use of antibodies to phosphothreonine. Methods Enzymol 201, 44–53.
Ogawa R, Ohtsuka M, Sasadaira H et al. (1985) Increase of phosphotyrosine-containing proteins in human carcinomas. Jpn J Cancer Res 76, 1049–1055.
Saper CB. (2005) An open letter to our readers on the use of antibodies. J Comp Neurol 493, 477–478.
Hsi ED. (2001) A practical approach for evaluating new antibodies in the clinical immunohistochemistry laboratory. Arch Pathol Lab Med 125, 289–294.
Yano T, Taura C, Shibata M et al. (1991) A monoclonal antibody to the phosphorylated form of glial fibrillary acidic protein: application to a non-radioactive method for measuring protein kinase activities. Biochem Biophys Res Commun 175, 1144–1151.
Czernik AJ, Girault JA, Nairn AC et al. (1991) Production of phosphorylation state-specific antibodies. Methods Enzymol 201, 264–283.
Krutzik PO, Irish JM, Nolan GP, Perez OD. (2004) Analysis of protein phosphorylation and cellular signaling events by flow cytometry: techniques and clinical applications. Clin Immunol 110, 206–221.
Espina V, Edmiston KH, Heiby M et al. (2008) A portrait of tissue phosphoprotein stability in the clinical tissue procurement process. Mol Cell Proteomics 7, 1998–2018.
Rikova K, Guo A, Zeng Q et al. (2007) Global survey of phosphotyrosine signaling identifies oncogenic kinases in lung cancer. Cell 131, 1190–1203.
Zheng H, Hu P, Quinn DF, Wang YK. (2005) Phosphotyrosine proteomic study of interferon alpha signaling pathway using a combination of immunoprecipitation and immobilized metal affinity chromatography. Mol Cell Proteomics 4, 721–730.
Shankaran H, Ippolito DL, Chrisler WB et al. (2009) Rapid and sustained nuclear-cytoplasmic ERK oscillations induced by epidermal growth factor. Mol Syst Biol 5, 332.
Bodo J, Durkin L, Hsi ED. (2009) Quantitative in situ detection of phosphoproteins in fixed tissues using quantum dot technology. J Histochem Cytochem 57, 701–708.
Fantin VR, Loboda A, Paweletz CP et al. (2008) Constitutive activation of signal transducers and activators of transcription predicts vorinostat resistance in cutaneous T-cell lymphoma. Cancer Res 68, 3785–3794.
Acknowledgments
The authors acknowledge Lisa Durkin for her technical expertise in immunostaining.
Author information
Authors and Affiliations
Corresponding author
Editor information
Editors and Affiliations
Rights and permissions
Copyright information
© 2011 Springer Science+Business Media, LLC
About this protocol
Cite this protocol
Bodo, J., Hsi, E.D. (2011). Selection and Validation of Antibodies for Signal Transduction Immunohistochemistry. In: Kalyuzhny, A. (eds) Signal Transduction Immunohistochemistry. Methods in Molecular Biology, vol 717. Humana Press. https://doi.org/10.1007/978-1-61779-024-9_2
Download citation
DOI: https://doi.org/10.1007/978-1-61779-024-9_2
Published:
Publisher Name: Humana Press
Print ISBN: 978-1-61779-023-2
Online ISBN: 978-1-61779-024-9
eBook Packages: Springer Protocols