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dc.contributor.authorGullaksen, Stein-Eriken_US
dc.contributor.authorBader, Luciusen_US
dc.contributor.authorHellesøy, Monicaen_US
dc.contributor.authorSulen, Andreen_US
dc.contributor.authorFagerholt, Oda Helen Ecken_US
dc.contributor.authorEngen, Caroline Benedicte Nitteren_US
dc.contributor.authorSkavland, Jørnen_US
dc.contributor.authorGjertsen, Bjørn Toreen_US
dc.contributor.authorGavasso, Soniaen_US
dc.PublishedGullaksen S, Bader L, Hellesøy M, Sulen A, Fagerholt OHE, Engen CB, Skavland J, Gjertsen BT, Gavasso S. Titrating complex mass cytometry panels. Cytometry Part A. 2019:95(7):792–796eng
dc.description.abstractWe describe here a simple and efficient antibody titration approach for cell‐surface markers and intracellular cell signaling targets for mass cytometry. The iterative approach builds upon a well‐characterized backbone panel of antibodies and analysis using bioinformatic tools such as SPADE. Healthy peripheral blood and bone marrow cells are stained with a pre‐optimized “backbone” antibody panel in addition to the progressively diluted (titrated) antibodies. Clustering based on the backbone panel enables the titration of each antibody against a rich hematopoietic background and assures that nonspecific binding and signal spillover can be quantified accurately. Using a slightly expanded backbone panel, antibodies quantifying changes in transcription factors and phosphorylated antigens are titrated on ex vivo stimulated cells to optimize sensitivity and evaluate baseline expression. Based on this information, complex panels of antibodies can be thoroughly optimized for use on healthy whole blood and bone marrow and are easily adaptable to the investigation of samples from for example clinical studies.en_US
dc.publisherWiley Periodicalseng
dc.rightsAttribution CC BY-NCeng
dc.subjectmass cytometryeng
dc.subjectpanel designeng
dc.subjectantibody titrationeng
dc.subjectwhole bloodeng
dc.subjectBone marroweng
dc.titleTitrating complex mass cytometry panelsen_US
dc.typePeer reviewed
dc.typeJournal article
dc.rights.holderCopyright 2019 The Author(s)
dc.source.journalCytometry Part A

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