Anti-DNA-RNA Hybrid [S9.6] Antibody

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

Highlights:

  • High specificity and affinity for DNA-RNA hybrids
  • Useful in the detection of R-loops
  • 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

DNA-RNA hybrids are a natural occurrence within eukaryotic cells, with levels of these hybrids increasing at sites with high transcriptional activity, such as during transcription initiation, repression, and elongation. Because RNA-DNA hybrids influence genomic instability, the S9.6 antibody is a useful reagent to help study the consequences of R-loops and lesions formed by these hybrids during DNA replication or other cellular processes. In addition, the S9.6 antibody is effective in recognizing RNA-DNA hybridization for microarray studies.

From the laboratory of Stephen H. Leppla, PhD, National Institute of Allergy and Infectious Diseases/NIH.

Catalog Number Product Size AVAILABILITY Price Qty
ENH001
Anti-DNA-RNA Hybrid [S9.6] Antibody, 100ug
100ug In stock
Price: $319.00
ENH002
Anti-DNA-RNA Hybrid [S9.6] Antibody, 1mg (10x100ug)
1mg (10x100ug) In stock
Price: $2,750.00
Specifications
Product Type: Antibody
Antigen: S9.6 ΦX174 bacteriophage-derived synthetic DNA/RNA antigen
Isotype: IgG2a
Clonality: Monoclonal
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
Species Immunized: BALB/c mouse
Paratope: DYYGSRWFDY (proposed)
Purification Method: Protein G
Buffer: 0.1M Sodium Phosphate, pH 7.4, 0.15M NaCl, 0.05% (w/v) Sodium Azide
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

Storage: -20C
Shipped: Cold packs
Provider
From the laboratory of Stephen H. Leppla, PhD, National Institute of Allergy and Infectious Diseases/NIH.
References

PDF Anti-DNA-RNA Hybrid [S9.6] Antibody - Publications by Application »

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