Chemogenetic detection and quantitation of H2O2 in living cells
Hydrogen peroxide (H2O2) is a natural product of aerobic metabolism. It acts as a signaling molecule and regulates fundamental cellular functions. However, it has remained difficult to measure intracellular H2O2 with high specificity and in a quantitative manner. Here, we present a detailed protocol...
Gespeichert in:
| Hauptverfasser: | , , |
|---|---|
| Dokumenttyp: | Article (Journal) |
| Sprache: | Englisch |
| Veröffentlicht: |
11 August 2025
|
| In: |
Nature protocols
Year: 2025, Pages: 1-24 |
| ISSN: | 1750-2799 |
| DOI: | 10.1038/s41596-025-01226-9 |
| Online-Zugang: | Verlag, lizenzpflichtig, Volltext: https://doi.org/10.1038/s41596-025-01226-9 Verlag, lizenzpflichtig, Volltext: https://www.nature.com/articles/s41596-025-01226-9 
|
| Verfasserangaben: | Mohammad Eid, Uladzimir Barayeu & Tobias P. Dick |
MARC
| LEADER | 00000naa a22000002c 4500 | ||
|---|---|---|---|
| 001 | 1946494577 | ||
| 003 | DE-627 | ||
| 005 | 20251218100956.0 | ||
| 007 | cr uuu---uuuuu | ||
| 008 | 251218s2025 xx |||||o 00| ||eng c | ||
| 024 | 7 | |a 10.1038/s41596-025-01226-9 |2 doi | |
| 035 | |a (DE-627)1946494577 | ||
| 035 | |a (DE-599)KXP1946494577 | ||
| 040 | |a DE-627 |b ger |c DE-627 |e rda | ||
| 041 | |a eng | ||
| 084 | |a 33 |2 sdnb | ||
| 100 | 1 | |a Eid, Mohammad |d 1993- |e VerfasserIn |0 (DE-588)1316133605 |0 (DE-627)1878214721 |4 aut | |
| 245 | 1 | 0 | |a Chemogenetic detection and quantitation of H2O2 in living cells |c Mohammad Eid, Uladzimir Barayeu & Tobias P. Dick |
| 246 | 3 | 3 | |a Chemogenetic detection and quantitation of H 2 O 2 in living cells |
| 264 | 1 | |c 11 August 2025 | |
| 300 | |b Illustrationen, Diagramme | ||
| 300 | |a 24 | ||
| 336 | |a Text |b txt |2 rdacontent | ||
| 337 | |a Computermedien |b c |2 rdamedia | ||
| 338 | |a Online-Ressource |b cr |2 rdacarrier | ||
| 500 | |a Im Text ist "2" jeweils tiefgestellt | ||
| 500 | |a Gesehen am 18.12.2025 | ||
| 520 | |a Hydrogen peroxide (H2O2) is a natural product of aerobic metabolism. It acts as a signaling molecule and regulates fundamental cellular functions. However, it has remained difficult to measure intracellular H2O2 with high specificity and in a quantitative manner. Here, we present a detailed protocol for a chemogenetic method that enables the detection and quantitation of H2O2 in living cells by converting intracellular H2O2 into fluorescent or luminescent signals. This is achieved by expressing the engineered heme peroxidase APEX2 in cells and subcellular locations of interest and by providing an appropriate fluorogenic or luminogenic substrate from outside. This method differs fundamentally from previously developed genetically encoded H2O2 probes; those are reversible and measure the balance between probe thiol oxidation and reduction. By contrast, APEX2 turns over its substrate irreversibly and therefore directly measures endogenous H2O2 availability. Our detailed step-by-step protocol covers the generation of APEX2-expressing cell lines, the implementation of fluorescent and luminescent measurements and examples for application. Ectopic expression of APEX2 can be achieved in 3 days, while the actual measurements typically require 1-2 h. This protocol is intended for entry-level scientists. | ||
| 650 | 4 | |a Chemical tools | |
| 650 | 4 | |a Sensors and probes | |
| 700 | 1 | |a Barayeu, Uladzimir |d 1993- |e VerfasserIn |0 (DE-588)1278905219 |0 (DE-627)1831805030 |4 aut | |
| 700 | 1 | |a Dick, Tobias P. |e VerfasserIn |0 (DE-588)1142415805 |0 (DE-627)100217807X |0 (DE-576)494813431 |4 aut | |
| 773 | 0 | 8 | |i Enthalten in |t Nature protocols |d Basingstoke : Nature Publishing Group, 2006 |g (2025), online ahead of print |h Online-Ressource |w (DE-627)515980552 |w (DE-600)2244966-8 |w (DE-576)276000455 |x 1750-2799 |7 nnas |a Chemogenetic detection and quantitation of H2O2 in living cells |
| 773 | 1 | 8 | |g year:2025 |g pages:1-24 |g extent:24 |a Chemogenetic detection and quantitation of H2O2 in living cells |
| 856 | 4 | 0 | |u https://doi.org/10.1038/s41596-025-01226-9 |x Verlag |x Resolving-System |z lizenzpflichtig |3 Volltext |7 1 |
| 856 | 4 | 0 | |u https://www.nature.com/articles/s41596-025-01226-9 |x Verlag |z lizenzpflichtig |3 Volltext |7 1 |
| 951 | |a AR | ||
| 992 | |a 20251218 | ||
| 993 | |a Article | ||
| 994 | |a 2025 | ||
| 998 | |g 1142415805 |a Dick, Tobias P. |m 1142415805:Dick, Tobias P. |p 3 |y j | ||
| 999 | |a KXP-PPN1946494577 |e 4829849975 | ||
| BIB | |a Y | ||
| SER | |a journal | ||
| JSO | |a {"relHost":[{"id":{"issn":["1750-2799"],"eki":["515980552"],"zdb":["2244966-8"]},"origin":[{"publisherPlace":"Basingstoke","dateIssuedDisp":"2006-","publisher":"Nature Publishing Group","dateIssuedKey":"2006"}],"physDesc":[{"extent":"Online-Ressource"}],"title":[{"title":"Nature protocols","title_sort":"Nature protocols"}],"recId":"515980552","language":["eng"],"type":{"media":"Online-Ressource","bibl":"periodical"},"disp":"Chemogenetic detection and quantitation of H2O2 in living cellsNature protocols","note":["Gesehen am 18. September 2018"],"part":{"pages":"1-24","year":"2025","extent":"24","text":"(2025), online ahead of print"},"titleAlt":[{"title":"Protocols"}],"pubHistory":["1.2006 -"]}],"physDesc":[{"extent":"24 S.","noteIll":"Illustrationen, Diagramme"}],"name":{"displayForm":["Mohammad Eid, Uladzimir Barayeu & Tobias P. Dick"]},"id":{"eki":["1946494577"],"doi":["10.1038/s41596-025-01226-9"]},"origin":[{"dateIssuedDisp":"11 August 2025","dateIssuedKey":"2025"}],"recId":"1946494577","language":["eng"],"type":{"bibl":"article-journal","media":"Online-Ressource"},"note":["Im Text ist \"2\" jeweils tiefgestellt","Gesehen am 18.12.2025"],"titleAlt":[{"title":"Chemogenetic detection and quantitation of H 2 O 2 in living cells"}],"person":[{"family":"Eid","given":"Mohammad","roleDisplay":"VerfasserIn","display":"Eid, Mohammad","role":"aut"},{"given":"Uladzimir","family":"Barayeu","role":"aut","roleDisplay":"VerfasserIn","display":"Barayeu, Uladzimir"},{"display":"Dick, Tobias P.","roleDisplay":"VerfasserIn","role":"aut","family":"Dick","given":"Tobias P."}],"title":[{"title_sort":"Chemogenetic detection and quantitation of H2O2 in living cells","title":"Chemogenetic detection and quantitation of H2O2 in living cells"}]} | ||
| SRT | |a EIDMOHAMMACHEMOGENET1120 | ||