High sensitivity immunolocalization of double and single-stranded DNA by a monoclonal antibody.
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.
Scheer
U
U
Messner
K
K
Hazan
R
R
Raska
I
I
Hansmann
P
P
Falk
H
H
Spiess
E
E
Franke
W W
WW
D000818:Animals; D000911:Antibodies, Monoclonal; D002460:Cell Line; D004247:DNA / Q000032:analysis*; D004277:DNA, Single-Stranded / Q000032:analysis*; D005455:Fluorescent Antibody Technique; D051379:Mice; D008854:Microscopy, Electron; D010999:Pleurodeles; D051381:Rats; D011919:Rats, Inbred Strains; D014319:Triturus; D014982:Xenopus laevis
D000911:Antibodies, Monoclonal; D004277:DNA, Single-Stranded; D004247:DNA