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Dictyostelium as a Model to Assess Site-Specific ADP-Ribosylation Events.

Methods in molecular biology (Clifton, N.J.) (2018-08-12)
Anna-Lena Kolb, Duen-Wei Hsu, Ana B A Wallis, Seiji Ura, Alina Rakhimova, Catherine J Pears, Nicholas D Lakin
ABSTRACT

The amoeba Dictyostelium discoideum is a single-cell organism that can undergo a simple developmental program, making it an excellent model to study the molecular mechanisms of cell motility, signal transduction, and cell-type differentiation. A variety of human genes that are absent or show limited conservation in other invertebrate models have been identified in this organism. This includes ADP-ribosyltransferases, also known as poly-ADP-ribose polymerases (PARPs), a family of proteins that catalyze the addition of single or poly-ADP-ribose moieties onto target proteins. The genetic tractability of Dictyostelium and its relatively simple genome structure makes it possible to disrupt PARP gene combinations, in addition to specific ADP-ribosylation sites at endogenous loci. Together, this makes Dictyostelium an attractive model to assess how ADP-ribosylation regulates a variety of cellular processes including DNA repair, transcription, and cell-type specification. Here we describe a range of techniques to study ADP-ribosylation in Dictyostelium, including analysis of ADP-ribosylation events in vitro and in vivo, in addition to approaches to assess the functional roles of this modification in vivo.

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Sigma-Aldrich
MONOCLONAL ANTI-HIS TAG antibody produced in mouse, clone 6AT18, IgG fraction of antiserum, buffered aqueous solution
Sigma-Aldrich
Anti-pan-ADP-ribose binding reagent, from Escherichia coli
Sigma-Aldrich
reagente anti-legame poli-ADP-ribosio, Anti-poly-ADP-ribose binding reagent is a reagent that selectively binds to ADP ribose for use in Western Blotting, Immunocytochemistry and Dot Blot.