Transfection Of Bone Marrow-Derived Mast Cells For Transcription Factor Luciferase Reporter Assays Protocol

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Title Transfection Of Bone Marrow-Derived Mast Cells For Transcription Factor Luciferase Reporter Assays
Source Nature Protocols: 10.1038/nprot.2006.218

Overview

Mast cells are now being understood as important intitiators and regulators in both innate and adaptive immunity in addition to their traditional roles in allergic and IgE-mediated immune responses.Transfection of primary leukocytes has traditionally been a challenging but much desired protocol. It allows not only the analysis of cells in a more natural state to a cell line system, it enables the direct comparison of, for example, transcriptional activity using luciferase reporters, in immune cells taken from genetically-altered mice. In addition, importantly it allows for "rescue experiments" in knockout cells as well as the ability to over-express or reconstitute wild-type and/or mutated constructs into wild-type or knockout cells for analysis of functional responses such as cytokine production, apoptosis, migration etc. By Kate Jeffrey Dr, Charles Mackay Prof and Tilman Brummer Dr

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