Abstract
The retinoblastoma protein (pRb) is an important tumor suppressor and cell cycle repressor. pRb is a phosphoprotein whose function is regulated primarily at the level of phosphorylation, and therefore, detecting pRb’s phosphorylation status in human tissue samples can be clinically informative. Unfortunately, detection of phosphorylated pRb residues can be technically challenging, as these residues can often be weak antigens. In this chapter, we describe an enhanced sensitivity immunohistochemistry protocol for the staining of phosphorylated serine 249 in pRb, in human lung tumor samples.
This is a preview of subscription content, log in via an institution.
Buying options
Tax calculation will be finalised at checkout
Purchases are for personal use only
Learn about institutional subscriptionsReferences
Harbour JW, Dean DC (2000) Rb function in cell-cycle regulation and apoptosis. Nat Cell Biol 2(4):E65–E67
Dyson N (1998) The regulation of E2F by pRB-family proteins. Genes Dev 12:2245–2262
Ross JF, Liu X, Dynlacht BD (1999) Mechanism of transcriptional repression of E2F by the retinoblastoma tumor suppressor protein. Mol Cell 3:195–205
Thomas DM, Carty SA, Piscopo DM, Lee J-S, Wang W-F et al (2001) The retinoblastoma protein acts as a transcriptional coactivator required for osteogenic differentiation. Mol Cell 8:303–316
Thomas DM, Yang H-S, Alexander K, Hinds PW (2003) Role of the retinoblastoma protein in differentiation and senescence. Cancer Biol Ther 2:2–8
Rubin SM (2013) Deciphering the retinoblastoma protein phosphorylation code. Trends Biochem Sci 38(1):12–19
Lundberg AS, Weinberg RA (1998) Functional inactivation of the retinoblastoma protein requires sequential modification by at least two distinct cyclin-cdk complexes. Mol Cell Biol 18(2):753–761
Buchkovich K, Duffy LA, Harlow E (1989) The retinoblastoma protein is phosphorylated during specific phases of the cell cycle. Cell 58(6):1097–1105
Burke JR, Liban TJ, Restrepo T, Lee HW, Rubin SM (2014) Multiple mechanisms for E2F binding inhibition by phosphorylation of the retinoblastoma protein C-terminal domain. J Mol Biol 426(1):245–255
Author information
Authors and Affiliations
Corresponding author
Editor information
Editors and Affiliations
Rights and permissions
Copyright information
© 2018 Springer Science+Business Media, LLC
About this protocol
Cite this protocol
Pérez-Morales, J., Núñez-Marrero, A., Santiago-Cardona, P.G. (2018). Immunohistochemical Detection of Retinoblastoma Protein Phosphorylation in Human Tumor Samples. In: Santiago-Cardona, P. (eds) The Retinoblastoma Protein. Methods in Molecular Biology, vol 1726. Humana Press, New York, NY. https://doi.org/10.1007/978-1-4939-7565-5_8
Download citation
DOI: https://doi.org/10.1007/978-1-4939-7565-5_8
Published:
Publisher Name: Humana Press, New York, NY
Print ISBN: 978-1-4939-7564-8
Online ISBN: 978-1-4939-7565-5
eBook Packages: Springer Protocols