Flow Cytometric Identification of Human IgE + B Lineage Subsets.
| Autor: | Pathmanandavel K; Garvan Institute of Medical Research, Darlinghurst, NSW, Australia. K.Pathmanandavel@garvan.org.au.; School of Clinical Medicine, Sydney, NSW, Australia. K.Pathmanandavel@garvan.org.au., Tangye SG; Garvan Institute of Medical Research, Darlinghurst, NSW, Australia.; School of Clinical Medicine, Sydney, NSW, Australia., Ma CS; Garvan Institute of Medical Research, Darlinghurst, NSW, Australia. c.ma@garvan.org.au.; School of Clinical Medicine, Sydney, NSW, Australia. c.ma@garvan.org.au. |
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| Jazyk: | angličtina |
| Zdroj: | Methods in molecular biology (Clifton, N.J.) [Methods Mol Biol] 2024; Vol. 2826, pp. 189-199. |
| DOI: | 10.1007/978-1-0716-3950-4_14 |
| Abstrakt: | The use of flow cytometry for immunophenotyping is contingent on the ability to accurately assign biological relevance to the detected signal. This process has historically been challenging when defining IgE expressing B cells or IgE expressing antibody-secreting cells due to widespread expression of receptors for IgE on various leukocyte subsets, including human B cells. Here we describe our implementation of intracellular staining for human IgE following a blocking step to negate the challenge of surface-bound IgE. We also describe our experience with a human B cell culture system that can be used to robustly validate this approach before application to primary human samples. Orthogonal confirmatory techniques remain essential; these are not described in detail, but several possible strategies are suggested. (© 2024. The Author(s), under exclusive license to Springer Science+Business Media, LLC, part of Springer Nature.) |
| Databáze: | MEDLINE |
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