RFA Baker's yeast replication factor A | Agrisera AS07 214 product information

product summary

company name :

Agrisera

product type :

antibody

product name :

RFA Baker's yeast replication factor A

catalog :

AS07 214

quantity :

50 µl

clonality :

polyclonal

host :

rabbit

conjugate :

nonconjugated

reactivity :

fission yeast, Saccharomyces cerevisiae S288c, baker's yeast

application :

western blot, immunoprecipitation, chromatin immunoprecipitation

more info or order :

Agrisera product webpage

citations: 7

Published Application/Species/Sample/Dilution	Reference
western blot; baker's yeast; fig 4d	Yeeles J, Deegan T, Janska A, Early A, Diffley J. Regulated eukaryotic DNA replication origin firing with purified proteins. Nature. 2015;519:431-5 pubmed publisher
western blot; fission yeast; fig 4	Lee L, Rodriguez J, Tsukiyama T. Chromatin remodeling factors Isw2 and Ino80 regulate checkpoint activity and chromatin structure in S phase. Genetics. 2015;199:1077-91 pubmed publisher
	He M, Liu J, Lu Y, Wu Z, Liu Y, Wu Z, et al. KEOPS complex promotes homologous recombination via DNA resection. Nucleic Acids Res. 2019;47:5684-5697 pubmed publisher
	Cheng X, Jobin Robitaille O, Billon P, Buisson R, Niu H, Lacoste N, et al. Phospho-dependent recruitment of the yeast NuA4 acetyltransferase complex by MRX at DNA breaks regulates RPA dynamics during resection. Proc Natl Acad Sci U S A. 2018;115:10028-10033 pubmed publisher
	Murayama Y, Samora C, Kurokawa Y, Iwasaki H, Uhlmann F. Establishment of DNA-DNA Interactions by the Cohesin Ring. Cell. 2018;172:465-477.e15 pubmed publisher
	Bantele S, Ferreira P, Gritenaite D, Boos D, Pfander B. Targeting of the Fun30 nucleosome remodeller by the Dpb11 scaffold facilitates cell cycle-regulated DNA end resection. elife. 2017;6: pubmed publisher
	Bentsen I, Nielsen I, Lisby M, Nielsen H, Gupta S, Mundbjerg K, et al. MRX protects fork integrity at protein-DNA barriers, and its absence causes checkpoint activation dependent on chromatin context. Nucleic Acids Res. 2013;41:3173-89 pubmed publisher

product information

Product number :

AS07 214

Product name :

RFA Baker's yeast replication factor A

Immunogen :

RPA from Saccharomyces cerevisiae consisting of three subunits RFA1 (70 kDa), RFA2 (30 kDa) and RFA3 (14 kDa); overexpressed in E.coli and purified by chromatography; no affinity tags were added to any of three subunits

Host :

Rabbit

Clonality :

Polyclonal

Purity :

Serum

Format :

Lyophilized

Quantity :

50 µl

Tested applications :

Immunoprecipitation (IP), Chromatin Immunoprecipitation (ChIP), Western blot (WB)

Recommended dilutions :

ChIP, 1 : 20 000 (WB)

Expected apparent MW :

70 + 30 + 14 kDa

Confirmed reactivity :

Saccharomyces cerevisiae

Predicted reactivity :

Saccharomyces cerevisiae

Not reactive in :

No confirmed exceptions from predicted reactivity are currently known.

Additional information :

Antibody was also successfully used in ChIP application Holstein et al. (2014).Load of 1 ng of the protein will allow to visualize two subunits of RPA, while load of 5 ng will allow to visualize all three subunits in Western blot technique.

Selected references :

Deshpande et al. (2017). Structural Basis of Mec1-Ddc2-RPA Assembly and Activation on Single-Stranded DNA at Sites of Damage. Mol Cell. 2017 Oct 19;68(2):431-445.e5. doi: 10.1016/j.molcel.2017.09.019. Chen et al. (2017). Dihydrocoumarin, an HDAC Inhibitor, Increases DNA Damage Sensitivity by Inhibiting Rad52. Int J Mol Sci. 2017 Dec 7;18(12). pii: E2655. doi: 10.3390/ijms18122655. Yeeles et al. (2015). Regulated eukaryotic DNA replication origin firing with purified proteins. Nature. 2015 Mar 4. doi: 10.1038/nature14285. Holstein et al. (2014). Interplay between Nonsense-Mediated mRNA Decay and DNA Damage Response Pathways Reveals that Stn1 and Ten1 Are the Key CST Telomere-Cap Components. Cell Rep. 2014 May 22;7(4):1259-69. doi: 10.1016/j.celrep.2014.04.017. Epub 2014 May 15. (ChIP)Deng et al. (2014). RPA antagonizes microhomology-mediated repair of DNA double-strand breaks. Nat Struct Mol Biol. 2014 Mar 9. doi: 10.1038/nsmb.2786. (western blot)Bentensen et al. (2013). MRX protects fork integrity at proteinâ€“DNA barriers, and its absence causes checkpoint activation dependent on chromatin contex. Nucleic Acids Res. 2013 Mar 1;41(5):3173-89. doi: 10.1093/nar/gkt051. (ChIP)

Pubmed ID's :

29033322, 29215575, 25739503, 24835988, 24608368, 23376930

Background :

Saccharomyces cerevisiaereplication protein A (RPA), also known as replication factor A (RFA) is a single-stranded DNA-binding protein that is required for multiple processes in eukaryotic DNA metabolism. Those processes include DNA replication, DNA repair, and recombination. Homologues to RPA have been identified in all eukaryotic organisms examined. RPA is heterotrimeric protein composed of subunits of approximately 70, 30, and 14 kDa. Members of this family bind nonspecifically to single-stranded DNA and interact with and/or modify the activities of multiple proteins. Alternative names: Replication protein A 69 kDa DNA-binding subunit, Single-stranded DNA-binding protein, DNA-binding protein BUF2, replication protein A 36 kDa subunit, DNA-binding protein BUF1 antibody

Reconstitution :

For reconstitution add 50 µl of sterile water

Storage :

Store lyophilized/reconstituted at -20°C; once reconstituted make aliquots to avoid repeated freeze-thaw cycles. Please, remember to spin tubes briefly prior to opening them to avoid any losses that might occur from lyophilized material adhering to the cap or sides of the tubes.

UniProt number :

P22336 , P26754

more info or order :

Agrisera product webpage