使用标记的 PPBD 肽定量人细胞系中多磷酸盐的方案。
Protocol to quantify polyphosphate in human cell lines using a tagged PPBD peptide.
机构信息
Basic Sciences Department, Universitat International de Catalunya, 08195 Barcelona, Spain.
出版信息
STAR Protoc. 2022 May 1;3(2):101363. doi: 10.1016/j.xpro.2022.101363. eCollection 2022 Jun 17.
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).
多聚磷酸盐（polyP）是一种进化上保守的磷酸盐聚合物，由于其浓度低和高不稳定性，在人类细胞中很难研究。首先，我们描述了如何表达和纯化 Xpress 标记的 PPBD（Ppx1 多聚磷酸盐结合结构域）。我们描述了使用 Xpress-PPBD、Xpress 抗体和 Alexa 缀合的二级抗体在 HEK293T 细胞中检测和定量核多聚磷酸盐。我们还在 SH-SY5Y、HeLa 和 HEK293 细胞中使用了该方案。有关使用和执行此方案的完整详细信息，请参阅 Samper-Martín 等人。（2021）。
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