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A protocol for measuring the activity of protein kinase C-delta in murine bone-marrow-derived dendritic cells.

ABSTRACT: Protein kinase C-δ (PKC-δ) is a key enzyme controlling growth, differentiation, and apoptosis in various cells, including immune cells. Here, we present a protocol to determine PKC-δ activation in response to increased membrane-bound diacylglycerol or phorbol-12-myristate-13-acetate treatment in murine bone-marrow-derived dendritic cells. We describe steps for dendritic cell differentiation, the isolation of plasma membrane lipids, and the quantification of diacylglycerol. We then detail procedures for measuring PKC-δ kinase activity by in vitro assay, indirect immunofluorescence, and western blotting experiments. For complete details on the use and execution of this protocol, please refer to Parsons et al.¹.

SUBMITTER: Mohati SM

PROVIDER: S-EPMC11338191 | biostudies-literature | 2024 Jul

REPOSITORIES: biostudies-literature

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A protocol for measuring the activity of protein kinase C-delta in murine bone-marrow-derived dendritic cells.

Mohati Seyedeh Mahdiye SM Matak Arash Mohammadi AM Makdissi Stephanie S Di Cara Francesca F

STAR protocols 20240726 3

Protein kinase C-δ (PKC-δ) is a key enzyme controlling growth, differentiation, and apoptosis in various cells, including immune cells. Here, we present a protocol to determine PKC-δ activation in response to increased membrane-bound diacylglycerol or phorbol-12-myristate-13-acetate treatment in murine bone-marrow-derived dendritic cells. We describe steps for dendritic cell differentiation, the isolation of plasma membrane lipids, and the quantification of diacylglycerol. We then detail procedu ...[more]

PMID: 39068659

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