Detection of Post-translationally Modified p53 by Western Blotting

Anna Estevan Barber; David W. Meek

doi:10.1007/978-1-0716-1217-0_2

Detection of Post-translationally Modified p53 by Western Blotting

Anna Estevan Barber, David W. Meek (Lead / Corresponding author)

Research output: Chapter in Book/Report/Conference proceeding › Chapter (peer-reviewed) › peer-review

Abstract

The p53 tumor suppressor has a central role in many key cellular processes including the DNA damage response, aging, stem cell differentiation, and fertility. p53 undergoes extensive regulatory post-translational modification through events such as phosphorylation, acetylation, methylation, and ubiquitylation. Here, we describe western blotting-based methodology for the detection and relative quantification of individual phosphorylation events in p53. While we focus on well-established N-terminal modifications for the purpose of illustration, this approach can be used to investigate other post-translational modifications of the protein, drawing upon a broad range of commercially available modification-specific antibodies.

Original language	English
Title of host publication	Cell Cycle Checkpoints
Subtitle of host publication	Methods and Protocols
Editors	James J. Manfredi
Place of Publication	New York
Publisher	Humana Press
Pages	7-18
Number of pages	12
Volume	2267
Edition	1
ISBN (Electronic)	9781071612170
ISBN (Print)	9781071612163 (hbk), 9781071612194 (pbk)
DOIs	https://doi.org/10.1007/978-1-0716-1217-0_2
Publication status	Published - 2021

Publication series

Name	Methods in Molecular Biology
Volume	2267
ISSN (Print)	1064-3745
ISSN (Electronic)	1940-6029

Keywords

DNA-damage response
Phospho-specific antibodies
Phosphorylation
Post-translational modifications
Western blot
p53

ASJC Scopus subject areas

Molecular Biology
Genetics

Access to Document

10.1007/978-1-0716-1217-0_2

Cite this

@inbook{5a8e230bd3cd468dadbe7f7e308c7f74,

title = "Detection of Post-translationally Modified p53 by Western Blotting",

abstract = "The p53 tumor suppressor has a central role in many key cellular processes including the DNA damage response, aging, stem cell differentiation, and fertility. p53 undergoes extensive regulatory post-translational modification through events such as phosphorylation, acetylation, methylation, and ubiquitylation. Here, we describe western blotting-based methodology for the detection and relative quantification of individual phosphorylation events in p53. While we focus on well-established N-terminal modifications for the purpose of illustration, this approach can be used to investigate other post-translational modifications of the protein, drawing upon a broad range of commercially available modification-specific antibodies.",

keywords = "DNA-damage response, Phospho-specific antibodies, Phosphorylation, Post-translational modifications, Western blot, p53",

author = "Barber, \{Anna Estevan\} and Meek, \{David W.\}",

note = "This work was supported by a studentship from Breast Cancer Now (UK), grant number: 2013NovPhD134.",

year = "2021",

doi = "10.1007/978-1-0716-1217-0\_2",

language = "English",

isbn = "9781071612163 (hbk)",

volume = "2267",

series = "Methods in Molecular Biology",

publisher = "Humana Press",

pages = "7--18",

editor = "Manfredi, \{James J.\}",

booktitle = "Cell Cycle Checkpoints",

address = "United States",

edition = "1",

}

Detection of Post-translationally Modified p53 by Western Blotting. / Barber, Anna Estevan; Meek, David W. (Lead / Corresponding author).
Cell Cycle Checkpoints: Methods and Protocols. ed. / James J. Manfredi. Vol. 2267 1. ed. New York: Humana Press, 2021. p. 7-18 (Methods in Molecular Biology; Vol. 2267).

Research output: Chapter in Book/Report/Conference proceeding › Chapter (peer-reviewed) › peer-review

TY - CHAP

T1 - Detection of Post-translationally Modified p53 by Western Blotting

AU - Barber, Anna Estevan

AU - Meek, David W.

N1 - This work was supported by a studentship from Breast Cancer Now (UK), grant number: 2013NovPhD134.

PY - 2021

Y1 - 2021

N2 - The p53 tumor suppressor has a central role in many key cellular processes including the DNA damage response, aging, stem cell differentiation, and fertility. p53 undergoes extensive regulatory post-translational modification through events such as phosphorylation, acetylation, methylation, and ubiquitylation. Here, we describe western blotting-based methodology for the detection and relative quantification of individual phosphorylation events in p53. While we focus on well-established N-terminal modifications for the purpose of illustration, this approach can be used to investigate other post-translational modifications of the protein, drawing upon a broad range of commercially available modification-specific antibodies.

AB - The p53 tumor suppressor has a central role in many key cellular processes including the DNA damage response, aging, stem cell differentiation, and fertility. p53 undergoes extensive regulatory post-translational modification through events such as phosphorylation, acetylation, methylation, and ubiquitylation. Here, we describe western blotting-based methodology for the detection and relative quantification of individual phosphorylation events in p53. While we focus on well-established N-terminal modifications for the purpose of illustration, this approach can be used to investigate other post-translational modifications of the protein, drawing upon a broad range of commercially available modification-specific antibodies.

KW - DNA-damage response

KW - Phospho-specific antibodies

KW - Phosphorylation

KW - Post-translational modifications

KW - Western blot

KW - p53

UR - http://www.scopus.com/inward/record.url?scp=85103555066&partnerID=8YFLogxK

U2 - 10.1007/978-1-0716-1217-0_2

DO - 10.1007/978-1-0716-1217-0_2

M3 - Chapter (peer-reviewed)

C2 - 33786782

SN - 9781071612163 (hbk)

SN - 9781071612194 (pbk)

VL - 2267

T3 - Methods in Molecular Biology

SP - 7

EP - 18

BT - Cell Cycle Checkpoints

A2 - Manfredi, James J.

PB - Humana Press

CY - New York

ER -

Detection of Post-translationally Modified p53 by Western Blotting

Abstract

Publication series

Keywords

ASJC Scopus subject areas

Access to Document

Other files and links

Fingerprint

Activation of "Gain-of-Function" Mutant p53 by Chemotherapeutic Drugs in Breast Cancer Progression: Mechanism and Implications for Choice of Treatment (PhD Studentship)

Cite this

Detection of Post-translationally Modified p53 by Western Blotting

Abstract

Publication series

Keywords

ASJC Scopus subject areas

Access to Document

Other files and links

Fingerprint

Projects

Activation of "Gain-of-Function" Mutant p53 by Chemotherapeutic Drugs in Breast Cancer Progression: Mechanism and Implications for Choice of Treatment (PhD Studentship)

Cite this