TY  - JOUR
A1  - Scheer, Ulrich
A1  - Messner, Karin
A1  - Hazan, Rachel
A1  - Raska, Ivan
A1  - Hansmann, Paul
A1  - Falk, Heinz
A1  - Spiess, Eberhard
A1  - Franke, Werner W.
T1  - High sensitivity immunolocalization of double and single-stranded DNA by a monoclonal antibody
N2  - A monoclonal antibody (AK 30-10) is described which specifically reacts with DNA both in double and single-stranded forms but not with other molecules and structures, including deoxyribonucleotides and RNAs. When used in immunocytochemical experiments on tissue sections and permeabilized cultured cells, this antibody detects DNA-containing structures, even when the DNA is present in very small amounts. Examples of high resolution detection include the DNA present in amplified extrachromosomal nucleoli, chromomeres of lampbrush chromosomes, mitochondria, chloroplasts and mycoplasmal particles. In immunoelectron microscopy using the immunogold technique, the DNA was localized in distinct substructures such as the "fibrillar centers" of nucleoli and certain stromal centers in chloroplasts. The antibody also reacts with DNA of chromatin of living cells, as shown by microinjection into cultured mitotic cells and into nuclei of amphibian oocytes. The potential value and the limitations of immunocytochemical DNA detection are discussed.
KW  - Cytologie
KW  - DNA antibodies
KW  - monoclonal antibodies
KW  - DNA immunolocalization
KW  - chromatin
KW  - mycoplasma tests
Y1  - 1987
U6  - http://nbn-resolving.de/urn/resolver.pl?urn:nbn:de:bvb:20-opus-41063
ER  - 
TY  - JOUR
A1  - Laine, Romain F.
A1  - Albecka, Anna
A1  - van de Linde, Sebastian
A1  - Rees, Eric J.
A1  - Crump, Colin M.
A1  - Kaminski, Clemens F.
T1  - Structural analysis of herpes simplex virus by optical super-resolution imaging
JF  - Nature Communications
N2  - Herpes simplex virus type-1 (HSV-1) is one of the most widespread pathogens among humans. Although the structure of HSV-1 has been extensively investigated, the precise organization of tegument and envelope proteins remains elusive. Here we use super-resolution imaging by direct stochastic optical reconstruction microscopy (dSTORM) in combination with a model-based analysis of single-molecule localization data, to determine the position of protein layers within virus particles. We resolve different protein layers within individual HSV-1 particles using multi-colour dSTORM imaging and discriminate envelope-anchored glycoproteins from tegument proteins, both in purified virions and in virions present in infected cells. Precise characterization of HSV-1 structure was achieved by particle averaging of purified viruses and model-based analysis of the radial distribution of the tegument proteins VP16, VP1/2 and pUL37, and envelope protein gD. From this data, we propose a model of the protein organization inside the tegument.
KW  - tegument protein pUL36
KW  - fluorescence microscopy
KW  - monoclonal antibodies
KW  - 3-dimensional structure
KW  - type-1
KW  - nuclear pore complex
KW  - reconstruction microscopy
KW  - localization microscopy
KW  - resolution
KW  - envelopment
Y1  - 2015
U6  - http://nbn-resolving.de/urn/resolver.pl?urn:nbn:de:bvb:20-opus-144623
VL  - 6
IS  - 5980
ER  - 
TY  - JOUR
A1  - Buchner, Erich
A1  - Blanco Redondo, Beatriz
A1  - Bunz, Melanie
A1  - Halder, Partho
A1  - Sadanandappa, Madhumala K.
A1  - Mühlbauer, Barbara
A1  - Erwin, Felix
A1  - Hofbauer, Alois
A1  - Rodrigues, Veronica
A1  - VijayRaghavan, K.
A1  - Ramaswami, Mani
A1  - Rieger, Dirk
A1  - Wegener, Christian
A1  - Förster, Charlotte
T1  - Identification and Structural Characterization of Interneurons of the Drosophila Brain by Monoclonal Antibodies of the Würzburg Hybridoma Library
JF  - PLoS ONE
N2  - Several novel synaptic proteins have been identified by monoclonal antibodies (mAbs) of the Würzburg hybridoma library generated against homogenized Drosophila brains, e.g. cysteine string protein, synapse-associated protein of 47 kDa, and Bruchpilot. However, at present no routine technique exists to identify the antigens of mAbs of our library that label only a small number of cells in the brain. Yet these antibodies can be used to reproducibly label and thereby identify these cells by immunohistochemical staining. Here we describe the staining patterns in the Drosophila brain for ten mAbs of the Würzburg hybridoma library. Besides revealing the neuroanatomical structure and distribution of ten different sets of cells we compare the staining patterns with those of antibodies against known antigens and GFP expression patterns driven by selected Gal4 lines employing regulatory sequences of neuronal genes. We present examples where our antibodies apparently stain the same cells in different Gal4 lines suggesting that the corresponding regulatory sequences can be exploited by the split-Gal4 technique for transgene expression exclusively in these cells. The detection of Gal4 expression in cells labeled by mAbs may also help in the identification of the antigens recognized by the antibodies which then in addition to their value for neuroanatomy will represent important tools for the characterization of the antigens. Implications and future strategies for the identification of the antigens are discussed.
KW  - cell staining
KW  - drosophila melanogaster
KW  - gene expression
KW  - hybridomas
KW  - immune serum
KW  - library screening
KW  - monoclonal antibodies
KW  - neurons
Y1  - 2013
U6  - http://nbn-resolving.de/urn/resolver.pl?urn:nbn:de:bvb:20-opus-97109
ER  -