Protocol to quantify polyphosphate in human cell lines using a tagged PPBD peptide
Author
Jiménez, Javier
Lázaro, Blanca
Sarrias, Ana
Tadeo, Francisco J.
Pérez-Montero, Marta
Clotet, Josep
Bru, Samuel
Publication date
2022ISSN
2666-1667
Abstract
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).
Document Type
Article
Document version
Published version
Language
English
Subject (CDU)
57 - Biological sciences in general
Keywords
Biologia cel·lular
Anticòs de microscòpia
Bioquímica de proteïnes
Expressió i purificació de proteïnes
Biología Celular
Anticuerpo de microscopía
Bioquímica de Proteínas
Expresión y purificación de proteínas
Cell Biology
Microscopy antibody
Protein biochemistry
Protein expression and purification
Pages
10
Publisher
Elsevier
Collection
3;2
Is part of
STAR Protocols
Citation
Jiménez, Javier; Lázaro, Blanca; Sarrias, Ana [et al.]. Protocol to quantify polyphosphate in human cell lines using a tagged PPBD peptide. STAR Protocols, 2022, 3(2), 101363. Disponible en: <https://www.sciencedirect.com/science/article/pii/S266616672200243X?via%3Dihub>. Fecha de acceso: 23 ene. 2024. DOI: 10.1016/j.xpro.2022.101363
This item appears in the following Collection(s)
- Ciències de la Salut [725]
Rights
Under a Creative Commons license.
Except where otherwise noted, this item's license is described as https://creativecommons.org/licenses/by/4.0/