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Kerafast Anti-DNA-RNA抗体ENH001简介

发布时间:2022-07-11   点击次数:81次

 

产品介绍:

Kerafast的货号:ENH001,Anti-DNA-RNA Hybrid [S9.6] Antibody,这种小鼠单克隆抗体是针对 ΦX174 噬菌体衍生的合成 DNA-RNA 抗原生成的,可识别各种长度的 RNA-DNA 杂合体。


特色:

可用于检测 R 环

对 DNA-RNA 杂交体的高特异性和亲和力

不与单链 DNA 或双链 DNA 发生交叉反应

对于富含 AU 的双链 RNA,观察到了轻微的交叉反应(约 5 倍以下)。

长度为 8、10、15 和 23 个碱基对的杂交体显示出高亲和力结合


DNA-RNA 杂合体是真核细胞中的一种自然现象,这些杂合体的水平在具有高转录活性的位点增加,例如在转录起始、抑制和延伸期间。由于 RNA-DNA 杂合体会影响基因组的不稳定性,因此 S9.6 抗体是一种有用的试剂,可帮助研究在 DNA 复制或其他细胞过程中由这些杂合体形成的 R 环和损伤的后果。此外,S9.6 抗体可有效识别用于微阵列研究的 RNA-DNA 杂交。

 

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


 


产品详情:

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).

 


参考文献:

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.

3. Lim J, Giri PK, Kazadi D, Laffleur B, Zhang W, Grinstein V, Pefanis E, Brown LM, Ladewig E, Martin O, Chen Y, Rabadan R, Boyer F, Rothschild G, Cogné M, Pinaud E, Deng H, Basu U. Nuclear Proximity of Mtr4 to RNA Exosome Restricts DNA Mutational Asymmetry. Cell. 2017 Apr 20;169(3):523-537.e15.

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.

5. De Cecco M, Ito T, Petrashen AP, Elias AE, Skvir NJ, Criscione SW, Caligiana A, Brocculi G, Adney EM, Boeke JD, Le O, Beauséjour C, Ambati J, Ambati K, Simon M, Seluanov A, Gorbunova V, Slagboom PE, Helfand SL, Neretti N, Sedivy JM. L1 drives IFN in senescent cells and promotes age-associated inflammation. Nature. 2019 Feb;566(7742):73-78.

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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