Isolation of the B Cell Immune Synapse for Proteomic Analysis

Diogo M. Cunha; Sara Hernández-Pérez; Pieta K. Mattila

doi:10.1007/978-1-0716-3135-5_25

Isolation of the B Cell Immune Synapse for Proteomic Analysis

Diogo M. Cunha, Sara Hernández-Pérez, Pieta K. Mattila^*

^*Korresponderande författare för detta arbete

Forskningsoutput: Kapitel i bok/konferenshandling › Kapitel › Vetenskaplig › Peer review

1 Citeringar (Scopus)

Sammanfattning

Recent technical developments have fueled increasing utilization of proteomics to gain new insights into various aspects of cellular behavior. In this chapter, we describe a method to specifically isolate immune synapses from mouse primary B cells. The method utilizes antibody-coated magnetic beads to induce the formation of the immune synapses and describes a protocol for the extraction of the cell-bead adhesions for mass spectrometry analysis. Finally, this method enables unveiling the large-scale protein content of the immune synapse.

Originalspråk	Engelska
Titel på värdpublikation	Methods in Molecular Biology
Förlag	Humana Press Inc.
Sidor	393-408
Antal sidor	16
Volym	2654
ISBN (tryckt)	978-1-0716-3134-8
DOI	https://doi.org/10.1007/978-1-0716-3135-5_25
Status	Publicerad - 28 apr. 2023
MoE-publikationstyp	A3 Del av bok eller annan forskningsbok

Publikationsserier

Namn	Methods in Molecular Biology
Volym	2654
ISSN (tryckt)	1064-3745
ISSN (elektroniskt)	1940-6029

Finansiering

We thank Dr. Alexandre Carisey (St Jude’s Research Hospital, Memphis, USA) for his generous help with setting up the protocol. We also acknowledge Prof. Jordan Orange and Dr. Anna Meyer for their pioneering efforts in the immune synapse isolation in NK cells (Department of Pediatrics, Vagelos College of Physicians and Surgeons, Columbia University Irving Medical Center, New York, New York, USA). Turku Proteomics Facility of Turku Bioscience and Biocenter Finland are acknowledged for providing the research infrastructures. This work was supported by the Academy of Finland (decision number: 339810 to PKM), Sigrid Juselius foundation (to PKM), Erasmus+ program (to DMC), the Finnish Cultural Foundation (Suomen Kulttuurirahasto; to SHP), and by InFLAMES Flagship Programme of the Academy of Finland (deci-sion number: 337530).

Finansiärer	Finansiärsnummer
Columbia University Irving Medical Center, New York, New York
Turku Proteomics Facility of Turku Bioscience
Department of Pediatrics, University of Florida
Vagelos College of Physicians and Surgeons, Columbia University
Academy of Finland	339810
Finska Kulturfonden	337530
Sigrid Jusélius Stiftelse
Biocenter Finland

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Isolation of the B Cell Immune Synapse for Proteomic Analysis

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