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A protocol for visualizing active cathepsin K in osteoclasts with a quenched-fluorescence-activity-based probe

2023
journal article
article
10.1016/j.xpro.2023.102465
Journal
STAR Protocols
20
Author
Korba Aleksandra

Ciastoń Izabela

Kozieł Joanna

Kasperkiewicz Paulina
Volume
4
Number
3
Article ID
102465
eISSN
2666-1667
Keywords in English

cell biology

cell-based assays

microscopy

molecular/chemical probes

Remarks

Bibliogr.

Language
English
Journal language
English
Abstract in English

Herein, we provide a protocol for visualizing active osteoclast cathepsin K (CatK) with the quenched-fluorescent-activity-based probe qTJK17. We describe steps for isolating peripheral blood mononuclear cells, their differentiation into osteoclasts, and TRAP staining using an acid phosphatase leukocyte kit. We then detail visualization of active CatK. The probe qTJK17 includes a reactive group, acyloxymethylketone, that binds to the CatK active site, recognition sequence, and fluorescence donor-acceptor pair. This protocol can determine the exact localization of active CatK in osteoclasts. For complete details on the use and execution of this protocol, please refer to Janiszewski et al. (2023).

Affiliation
Wydział Biochemii, Biofizyki i Biotechnologii : Zakład Mikrobiologii
cris.lastimport.wos2024-04-10T00:42:52Z
dc.abstract.enHerein, we provide a protocol for visualizing active osteoclast cathepsin K (CatK) with the quenched-fluorescent-activity-based probe qTJK17. We describe steps for isolating peripheral blood mononuclear cells, their differentiation into osteoclasts, and TRAP staining using an acid phosphatase leukocyte kit. We then detail visualization of active CatK. The probe qTJK17 includes a reactive group, acyloxymethylketone, that binds to the CatK active site, recognition sequence, and fluorescence donor-acceptor pair. This protocol can determine the exact localization of active CatK in osteoclasts. For complete details on the use and execution of this protocol, please refer to Janiszewski et al. (2023).pl
dc.affiliationWydział Biochemii, Biofizyki i Biotechnologii : Zakład Mikrobiologiipl
dc.contributor.authorKorba, Aleksandrapl
dc.contributor.authorCiastoń, Izabela - 163063 pl
dc.contributor.authorKozieł, Joanna - 129350 pl
dc.contributor.authorKasperkiewicz, Paulinapl
dc.date.accessioned2023-09-25T12:30:34Z
dc.date.available2023-09-25T12:30:34Z
dc.date.issued2023pl
dc.date.openaccess0
dc.description.accesstimew momencie opublikowania
dc.description.additionalBibliogr.pl
dc.description.number3pl
dc.description.versionostateczna wersja wydawcy
dc.description.volume4pl
dc.identifier.articleid102465pl
dc.identifier.doi10.1016/j.xpro.2023.102465pl
dc.identifier.eissn2666-1667pl
dc.identifier.urihttps://ruj.uj.edu.pl/xmlui/handle/item/319708
dc.languageengpl
dc.language.containerengpl
dc.rightsUdzielam licencji. Uznanie autorstwa - Użycie niekomercyjne - Bez utworów zależnych 4.0 Międzynarodowa*
dc.rights.licenceCC-BY-NC-ND
dc.rights.urihttp://creativecommons.org/licenses/by-nc-nd/4.0/legalcode.pl*
dc.share.typeotwarte czasopismo
dc.subject.encell biologypl
dc.subject.encell-based assayspl
dc.subject.enmicroscopypl
dc.subject.enmolecular/chemical probespl
dc.subtypeArticlepl
dc.titleA protocol for visualizing active cathepsin K in osteoclasts with a quenched-fluorescence-activity-based probepl
dc.title.journalSTAR Protocolspl
dc.typeJournalArticlepl
dspace.entity.typePublication
cris.lastimport.wos
2024-04-10T00:42:52Z
dc.abstract.enpl
Herein, we provide a protocol for visualizing active osteoclast cathepsin K (CatK) with the quenched-fluorescent-activity-based probe qTJK17. We describe steps for isolating peripheral blood mononuclear cells, their differentiation into osteoclasts, and TRAP staining using an acid phosphatase leukocyte kit. We then detail visualization of active CatK. The probe qTJK17 includes a reactive group, acyloxymethylketone, that binds to the CatK active site, recognition sequence, and fluorescence donor-acceptor pair. This protocol can determine the exact localization of active CatK in osteoclasts. For complete details on the use and execution of this protocol, please refer to Janiszewski et al. (2023).
dc.affiliationpl
Wydział Biochemii, Biofizyki i Biotechnologii : Zakład Mikrobiologii
dc.contributor.authorpl
Korba, Aleksandra
dc.contributor.authorpl
Ciastoń, Izabela - 163063
dc.contributor.authorpl
Kozieł, Joanna - 129350
dc.contributor.authorpl
Kasperkiewicz, Paulina
dc.date.accessioned
2023-09-25T12:30:34Z
dc.date.available
2023-09-25T12:30:34Z
dc.date.issuedpl
2023
dc.date.openaccess
0
dc.description.accesstime
w momencie opublikowania
dc.description.additionalpl
Bibliogr.
dc.description.numberpl
3
dc.description.version
ostateczna wersja wydawcy
dc.description.volumepl
4
dc.identifier.articleidpl
102465
dc.identifier.doipl
10.1016/j.xpro.2023.102465
dc.identifier.eissnpl
2666-1667
dc.identifier.uri
https://ruj.uj.edu.pl/xmlui/handle/item/319708
dc.languagepl
eng
dc.language.containerpl
eng
dc.rights*
Udzielam licencji. Uznanie autorstwa - Użycie niekomercyjne - Bez utworów zależnych 4.0 Międzynarodowa
dc.rights.licence
CC-BY-NC-ND
dc.rights.uri*
http://creativecommons.org/licenses/by-nc-nd/4.0/legalcode.pl
dc.share.type
otwarte czasopismo
dc.subject.enpl
cell biology
dc.subject.enpl
cell-based assays
dc.subject.enpl
microscopy
dc.subject.enpl
molecular/chemical probes
dc.subtypepl
Article
dc.titlepl
A protocol for visualizing active cathepsin K in osteoclasts with a quenched-fluorescence-activity-based probe
dc.title.journalpl
STAR Protocols
dc.typepl
JournalArticle
dspace.entity.type
Publication
Affiliations
Wydział Biochemii, Biofizyki i Biotechnologii
Ciastoń, Izabela
Kozieł, Joanna
No affiliation
Korba, Aleksandra
Kasperkiewicz, Paulina

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