Lead to changes in RNA base-pairing or RNA folding ( Motorin and Helm 2011 Polikanov et al. Bases can be chemically modified to gain or lose specific biophysical properties. Soon after the discovery of RNAs with noncoding functions such as transfer RNAs (tRNAs) or ribosomal RNAs (rRNAs), it becameĮvident that RNAs can be heavily modified and these modifications are important for their structures and functions ( Littlefield and Dunn 1958 Bergquist and Matthews 1962). RNA molecules are composed of nucleotides carrying the four bases adenine (A), guanine (G), cytosine (C), and uracil (U). Previous Section Next Section INTRODUCTION Here, we provide an experimental toolbox that allows for testing antibody performance prior to their use.

Therefore, it is critical that the affinity and specificity is rigorously tested using complementaryĪpproaches. The low abundance of the modification, particularly on mRNAs, generated data sets might resemble noise rather than specific Moreover, they are not always well characterized and due to Used but have limitations such as low affinity or specificity. Antibodies targeting modified nucleotides are often Well characterized, their physiological relevance remains largely elusive. However, since their abundances are low and tools used for their corroboration are often not Indeed, modifications including m 1A, m 5C, m 7G, 2′-OMe, and Ψ were detected. M 6A on mRNA and its important role in gene regulation stimulated the field to investigate whether additional modifications are Corresponding author: Ĭhemical modifications are found on almost all RNAs and affect their coding and noncoding functions.10Research Unit Molecular Immune Regulation, Helmholtz Zentrum München, Deutsches Forschungszentrum für Gesundheit und Umwelt.Internal Medicine III, University Hospital Heidelberg, 69120 Heidelberg, Germany 9Bioinformatics and Systems Cardiology, Klaus Tschira Institute for Integrative Computational Cardiology and Department of.8Institute of Molecular Medicine, Section for Molecular Cell Biology, Faculty of Medicine, Martin Luther University Halle-Wittenberg,Ĭharles Tanford Protein Centre, 06120 Halle, Germany.7Institute of Molecular Biology (IMB), Ackermannweg 4, 55128 Mainz, Germany.6Gene Center, Ludwig-Maximilians-University Munich, 81377 Munich, Germany.5Cell Biology and Epigenetics, Technische Universität Darmstadt, 64287 Darmstadt, Germany.4Institute of Pharmacy and Biochemistry, Johannes-Gutenberg-Universität Mainz, 55128 Mainz, Germany.3Institute for Immunology, Biomedical Center of the Ludwig-Maximilians-University München, 82152 Planegg-Martinsried, Germany.Research Center for Environmental Health (GmbH), 85764 Neuherberg, Germany 2Institute for Diabetes and Obesity, Monoclonal Antibody Core Facility and Research Group, Helmholtz Zentrum München, German.1Regensburg Center for Biochemistry, Laboratory for RNA Biology, University of Regensburg, 93053 Regensburg, Germany.

Antibodies in northern blot analysis