Anti-DNA-RNA hybrid [S9.6]

Absolute Antibody

Anti-DNA-RNA hybrid [S9.6]

7

DNA-RNA hybrids naturally occur in eukaryotic cells, with their levels increasing in regions of high transcriptional activity, such as during transcription initiation, repression, and elongation. These hybrids play a crucial role in influencing genomic instability, making the anti-DNA-RNA hybrid (S9.6) antibody an invaluable tool for studying R-loops and hybrid-induced lesions during DNA replication and other cellular processes. This antibody also excels in recognizing RNA-DNA hybridization in microarray studies. Key applications of the S9.6 antibody include affinity binding assays (ABA), chromatin immunoprecipitation (ChIP), chromatin immunoprecipitation-sequencing (ChIP-seq), dot blot (DB), fluorescent in situ hybridization (FISH), electrophoretic mobility shift assays (EMSA), surface plasmon resonance (SPR), immunofluorescence (IF), immunocytochemistry (ICC), and immunoprecipitation (IP). For example, in ABA, this antibody binds DNA-RNA heteropolymers and poly(I)-poly(dC) equally, but requires 100-fold higher levels of poly(A)-poly(dT) to achieve similar binding levels (Boguslawski et al., 1986; PMID: 2422282). In a ChIP analysis, a representative lot detected elevated DNA-RNA hybrid levels at four actively transcribed genes following shRNA-mediated knockdown of BRCA1 or BRCA2, but not PCID2 or RAD51 in HeLa cells (Bhatia et al., 2014; PMID: 24896180). Similarly, in ChIP-seq, the antibody detected genome-wide distribution of DNA-RNA hybrids in budding yeast (El Hage et al., 2014; PMID: 25357144). In ICC and IP analyses, the antibody immunoprecipitated in vitro transcribed R-loop substrates (DNA-RNA hybrids) but not double-stranded DNA (dsDNA) (Ginno et al., 2012; PMID: 22387027). Additionally, this antibody has been used for IF and proximity ligation assays (PLA) to detect DNA-RNA hybrids. In IF, it revealed DNA-RNA hybrid accumulation in HeLa cells during S/G2/M phases following ATAD5 depletion. In PLA, it assessed the proximity between DNA-RNA hybrids and PCNA (Kim et al., 2020; PMID: 32542338). Moreover, the Fab version of this antibody demonstrated binding to a 13-bp DNA/RNA hybrid, which was further studied via X-ray crystallography to examine its binding structure (Bou-Nader et al., 2022; PMID: 35347133).

Dot Blot, ImmunoFluorescence, ImmunoPrecipitation, ChIP Chromatin ImmunoPrecipitation, ImmunoCytoChemistry, Other Application

Research Use Only

This chimeric goat antibody was made using the variable domain sequences of the original Mouse IgG2a format, for improved compatibility with existing reagents, assays and techniques.

-20°C,2°C to 8°C

12352203