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University of Nebraska Medical Center

Journal Articles: Surgery

Apoptosis

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Full-Text Articles in Medicine and Health Sciences

Anti-Human Interleukin(Il)-4 Clone 8d4-8 Cross-Reacts With Myosin-9 Associated With Apoptotic Cells And Should Not Be Used For Flow Cytometry Applications Querying Il-4 Expression, Robert Z. Harms, Kiana Borengasser, Vikas Kumar, Nora Sarvetnick Jan 2019

Anti-Human Interleukin(Il)-4 Clone 8d4-8 Cross-Reacts With Myosin-9 Associated With Apoptotic Cells And Should Not Be Used For Flow Cytometry Applications Querying Il-4 Expression, Robert Z. Harms, Kiana Borengasser, Vikas Kumar, Nora Sarvetnick

Journal Articles: Surgery

Interleukin(IL)-4 is produced by T cells and other leukocytes and is a critical mediator of monocyte and B cell responses. During routine flow cytometry panel validation for the investigation of intracellular cytokines, we observed unique IL-4 expression patterns associated with the widely available monoclonal antibody 8D4-8. Namely, IL-4 (8D4-8) expression was observed in the absence of cellular activation and enhanced following staurosporine exposure. Mass spectrometry analysis of immunoprecipitates from peripheral blood lymphocytes (PBL) revealed that 8D4-8 cross-reacts with the ubiquitous cytoskeletal protein myosin-9. We confirmed these results by western blotting immunoprecipitates, using immunofluorescence among staurosporine-treated Caco-2 cells, and by surface-labeling …