產(chǎn)品介紹：

Kerafast的貨號：ENH001，Anti-DNA-RNA Hybrid [S9.6] Antibody，這種小鼠單克隆抗體是針對 ΦX174 噬菌體衍生的合成 DNA-RNA 抗原生成的，可識別各種長度的 RNA-DNA 雜合體。

特色：

可用于檢測 R 環(huán)

對 DNA-RNA 雜交體的高特異性和親和力

不與單鏈 DNA 或雙鏈 DNA 發(fā)生交叉反應(yīng)

對于富含 AU 的雙鏈 RNA，觀察到了輕微的交叉反應(yīng)（約 5 倍以下）。

長度為 8、10、15 和 23 個堿基對的雜交體顯示出高親和力結(jié)合

DNA-RNA 雜合體是真核細胞中的一種自然現(xiàn)象，這些雜合體的水平在具有高轉(zhuǎn)錄活性的位點增加，例如在轉(zhuǎn)錄起始、抑制和延伸期間。由于 RNA-DNA 雜合體會影響基因組的不穩(wěn)定性，因此 S9.6 抗體是一種有用的試劑，可幫助研究在 DNA 復(fù)制或其他細胞過程中由這些雜合體形成的 R 環(huán)和損傷的后果。此外，S9.6 抗體可有效識別用于微陣列研究的 RNA-DNA 雜交。

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

 

產(chǎn)品詳情：

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).

 

參考文獻：

1. Dutrow N, Nix DA, Holt D, Milash B, Dalley B, Westbroek E, Parnell TJ, Cairns BR. Dynamic transcriptome of Schizosaccharomyces pombe shown by RNA-DNA hybrid mapping. Nat Genet. 2008 Aug;40(8):977-86.

2. Bhatia V, Barroso SI, García-Rubio ML, Tumini E, Herrera-Moyano E, Aguilera A. BRCA2 prevents R-loop accumulation and associates with TREX-2 mRNA export factor PCID2. Nature. 2014 Jul 17;511(7509):362-5.

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4. Lang KS, Hall AN, Merrikh CN, Ragheb M, Tabakh H, Pollock AJ, Woodward JJ, Dreifus JE, Merrikh H. Replication-Transcription Conflicts Generate R-Loops that Orchestrate Bacterial Stress Survival and Pathogenesis. Cell. 2017 Aug 10;170(4):787-799.e18.

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6. Herold S, Kalb J, Büchel G, Ade CP, Baluapuri A, Xu J, Koster J, Solvie D, Carstensen A, Klotz C, Rodewald S, Schülein-Völk C, Dobbelstein M, Wolf E, Molenaar J, Versteeg R, Walz S, Eilers M. Recruitment of BRCA1 limits MYCN-driven accumulation of stalled RNA polymerase. Nature. 2019 Mar;567(7749):545-549

7. Sanz LA, Chédin F. High-resolution, strand-specific R-loop mapping via S9.6-based DNA-RNA immunoprecipitation and high-throughput sequencing. Nat Protoc. 2019 Jun;14(6):1734-1755.

8. Graf M, Bonetti D, Lockhart A, Serhal K, Kellner V, Maicher A, Jolivet P, Teixeira MT, Luke B. Telomere Length Determines TERRA and R-Loop Regulation through the Cell Cycle. Cell. 2017 Jun 29;170(1):72-85.e14.

9. Gorthi A, Romero JC, Loranc E, Cao L, Lawrence LA, Goodale E, Iniguez AB, Bernard X, Masamsetti VP, Roston S, Lawlor ER, Toretsky JA, Stegmaier K, Lessnick SL, Chen Y, Bishop AJR. EWS-FLI1 increases transcription to cause R-loops and block BRCA1 repair in Ewing sarcoma. Nature. 2018 Mar 15;555(7696):387-391.

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