This mouse monoclonal antibody was generated against a ΦX174 bacteriophage-derived synthetic DNA–RNA antigen and recognizes RNA-DNA hybrids of various lengths.

Highlights:

• Useful in the detection of R-loops
• High specificity and affinity for DNA-RNA hybrids
• Does NOT cross-react with single-stranded DNA or double-stranded DNA
• Minor cross-reaction (~5-fold less) has been observed for AU-rich double-stranded RNA.
• High affinity binding shown for hybrids of 8, 10, 15, and 23 base pairs in length

Product Type:	Antibody
Name:	Anti-DNA-RNA Hybrid [S9.6]
Antigen:	S9.6 ΦX174 bacteriophage-derived synthetic DNA–RNA antigen
Isotype:	Rabbit IgG
Fusion Tag(s):	Mouse Fab version contains His-tag
Clone Name:	S9.6
Reactivity:	High specificity and affinity for DNA/RNA hybrids and other A-form nucleic acid hybrids
Immunogen:	ΦX174 bacteriophage-derived synthetic DNA/RNA
Purification Method:	Protein A/G
Buffer:	ENHOO1: PBS, 0.05% (w/v) Sodium Azide Ab01137- : PBS with 0.02% Proclin 300
Tested Applications:	Dot Blot Analysis: 0.2 µg/mL. Affinity Binding Assay: Clone S9.6 bound the DNA-RNA heteropolymer and poly(I)-poly(dC) equally, but 100-fold higher levels of poly(A)-poly(dT) were required to achieve a similar degree of binding. Single-stranded DNA, double-stranded DNA and RNA, and ribosomal RNA were not bound by clone S9.6 (Boguslawski, S.J., et al. (1986). J. Immunol Methods. 89(1):123-130). Chromatin Immunoprecipitation (ChIP) Analysis: A representative lot detected increased DNA RNA hybrids at four actively transcribed genes upon shRNA-mediated knockdown of BRCA1 or BRCA2, but not PCID2 or RAD51 in HeLa cells (Bhatia, V., et al. (2014). Nature. 511(7509):362-365). Chromatin Immunoprecipitation (ChIP) Analysis: A representative lot detected R-loops formed over beta-actin gene using HeLa chromatin preparation. RNase H treatment of the chromatin preparation prevented clone S9.6 from immunoprecipitating target chromatin fragments (Skourti-Stathaki, K., et al. (2011). Mol. Cell. 42(6):794-805). Chromatin Immunoprecipitation-sequencing (ChIP-seq) Analysis: A representative lot detected genome-wide distribution of DNA-RNA hybrids in budding yeast by ChIP-seq analysis (El Hage, A., et al. (2014). PLoS Genet. 10(10):e1004716). Immunocytochemistry Analysis: Representative lots immunolocalized nuclear R loops by fluorescent immunocytochemistry staining of methanol-fixed H1 human embryonic stem cells (hESCs) and formaldehyde-fixed HeLa cells (Bhatia, V., et al. (2014). Nature. 511(7509):362-365; Ginno, P.A., et al. (2012). Mol. Cell. 45(6):814-825). Immunoprecipitation Analysis: A representative lot immunoprecipitated in vitro transcribed R-loop substrate (DNA-RNA hybrid), but not doouble-stranded DNA (dsDNA) (Ginno, P.A., et al. (2012). Mol. Cell. 45(6):814-825). See also: S9.6 Publications by Application