Protocol to quantify polyphosphate in human cell lines using a tagged PPBD peptide

Polyphosphate (polyP) is an evolutionarily conserved polymer of phosphates that is difficult to study in human cells because of its low concentration and high lability. First, we described how to express and purify Xpress-tagged PPBD (Ppx1 PolyP Binding Domain). We describe the detection and quantif...

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Autores principales: Jiménez, Javier, Lázaro, Blanca, Sarrias, Ana, Tadeo, Francisco J., Pérez-Montero, Marta, Clotet, Josep, Bru, Samuel
Formato: Online Artículo Texto
Lenguaje:English
Publicado: Elsevier 2022
Materias:
Protocol
Acceso en línea:https://www.ncbi.nlm.nih.gov/pmc/articles/PMC9079103/
https://www.ncbi.nlm.nih.gov/pubmed/35542178
http://dx.doi.org/10.1016/j.xpro.2022.101363
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author Jiménez, Javier
Lázaro, Blanca
Sarrias, Ana
Tadeo, Francisco J.
Pérez-Montero, Marta
Clotet, Josep
Bru, Samuel
author_facet Jiménez, Javier
Lázaro, Blanca
Sarrias, Ana
Tadeo, Francisco J.
Pérez-Montero, Marta
Clotet, Josep
Bru, Samuel
author_sort Jiménez, Javier
collection PubMed
description Polyphosphate (polyP) is an evolutionarily conserved polymer of phosphates that is difficult to study in human cells because of its low concentration and high lability. First, we described how to express and purify Xpress-tagged PPBD (Ppx1 PolyP Binding Domain). We describe the detection and quantification of nuclear polyP in HEK293T cells using Xpress-PPBD, Xpress antibody, and Alexa-conjugated secondary antibodies. We have also used this protocol in SH-SY5Y HeLa and HEK293 cells. For complete details on the use and execution of this protocol, please refer to Samper-Martín et al. (2021).
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spelling pubmed-90791032022-05-09 Protocol to quantify polyphosphate in human cell lines using a tagged PPBD peptide Jiménez, Javier Lázaro, Blanca Sarrias, Ana Tadeo, Francisco J. Pérez-Montero, Marta Clotet, Josep Bru, Samuel STAR Protoc Protocol Polyphosphate (polyP) is an evolutionarily conserved polymer of phosphates that is difficult to study in human cells because of its low concentration and high lability. First, we described how to express and purify Xpress-tagged PPBD (Ppx1 PolyP Binding Domain). We describe the detection and quantification of nuclear polyP in HEK293T cells using Xpress-PPBD, Xpress antibody, and Alexa-conjugated secondary antibodies. We have also used this protocol in SH-SY5Y HeLa and HEK293 cells. For complete details on the use and execution of this protocol, please refer to Samper-Martín et al. (2021). Elsevier 2022-05-01 /pmc/articles/PMC9079103/ /pubmed/35542178 http://dx.doi.org/10.1016/j.xpro.2022.101363 Text en © 2022 The Author(s) https://creativecommons.org/licenses/by/4.0/This is an open access article under the CC BY license (http://creativecommons.org/licenses/by/4.0/).
spellingShingle Protocol
Jiménez, Javier
Lázaro, Blanca
Sarrias, Ana
Tadeo, Francisco J.
Pérez-Montero, Marta
Clotet, Josep
Bru, Samuel
Protocol to quantify polyphosphate in human cell lines using a tagged PPBD peptide
title Protocol to quantify polyphosphate in human cell lines using a tagged PPBD peptide
title_full Protocol to quantify polyphosphate in human cell lines using a tagged PPBD peptide
title_fullStr Protocol to quantify polyphosphate in human cell lines using a tagged PPBD peptide
title_full_unstemmed Protocol to quantify polyphosphate in human cell lines using a tagged PPBD peptide
title_short Protocol to quantify polyphosphate in human cell lines using a tagged PPBD peptide
title_sort protocol to quantify polyphosphate in human cell lines using a tagged ppbd peptide
topic Protocol
url https://www.ncbi.nlm.nih.gov/pmc/articles/PMC9079103/
https://www.ncbi.nlm.nih.gov/pubmed/35542178
http://dx.doi.org/10.1016/j.xpro.2022.101363
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