產品介(jie)紹：

Kerafast的(de)貨(huo)號：ENH001，Anti-DNA-RNA Hybrid [S9.6] Antibody，這種小鼠(shu)單克隆抗體(ti)是針對 ΦX174 噬(shi)菌體(ti)衍(yan)生(sheng)(sheng)的(de)合(he)(he)成(cheng) DNA-RNA 抗原生(sheng)(sheng)成(cheng)的(de)，可識別各種長度的(de) RNA-DNA 雜(za)合(he)(he)體(ti)。

特色：

可用于檢測 R 環

對 DNA-RNA 雜交(jiao)體的高特異性和親和力

不與(yu)單鏈(lian) DNA 或雙(shuang)鏈(lian) DNA 發(fa)生交叉反應

對(dui)于富(fu)含 AU 的雙鏈(lian) RNA，觀察到(dao)了輕微的交叉(cha)反應（約 5 倍以下）。

長度為 8、10、15 和(he) 23 個堿基(ji)對的雜(za)交體顯示出(chu)高親(qin)和(he)力結合(he)

DNA-RNA 雜(za)合體是真核細胞(bao)中的(de)一(yi)(yi)種(zhong)自(zi)然(ran)現象，這些雜(za)合體的(de)水平在具有(you)高轉錄(lu)活性的(de)位(wei)點增(zeng)加，例如在轉錄(lu)起始、抑制(zhi)和(he)延伸(shen)期間(jian)。由(you)于(yu) RNA-DNA 雜(za)合體會影響(xiang)基(ji)因組的(de)不穩定性，因此 S9.6 抗體是一(yi)(yi)種(zhong)有(you)用(yong)的(de)試劑，可(ke)幫助研究(jiu)在 DNA 復制(zhi)或(huo)其他細胞(bao)過(guo)程(cheng)中由(you)這些雜(za)合體形成的(de) R 環和(he)損傷的(de)后果。此外，S9.6 抗體可(ke)有(you)效識別用(yong)于(yu)微陣列研究(jiu)的(de) RNA-DNA 雜(za)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

 

產品詳(xiang)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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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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