PAR/pADPr Antibody Summary
Applications
Please Note: Optimal dilutions should be determined by each laboratory for each application. General Protocols are available in the Technical Information section on our website.
Scientific Data
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Detection of PAR/pADPr by Western Blot. Western blot analysis of Wehi cells untreated (H) and treated (T) with 25 μM Etoposide for 4 hours at 37 °C. Cells were lysed in Tris-Glycine SDS sample buffer at the concentration of 1 x 107cells/ml, and 10 μl of clarified lysate were loaded per well of 4-20% Tris-Glycine gel. Proteins were transferred onto an Immobilon FL membrane and ribosylated proteins were detected using Trevigen's polyclonal anti-PAR antibody (cat# 4336-APC-050) followed by an IR680-conjugated secondary antibody (Licor). The membrane was scanned using an Odyssey Infrared Imaging System (Licor).
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Preparation and Storage
Background: PAR/pADPr
Procedure for Immunoblotting using Peroxidase Detection:
Blotting buffer: 12 mM Tris base, 96 mM Glycine, and 15% MeOH.
Blocking solution: 5% (w/v) nonfat dry milk in TBS-0.1% Tween.
Antibody solution: 5% (w/v) nonfat dry milk, in TBS-0.1% Tween.
Transfer the electrophoresed proteins to a PVDF membrane and incubate the membrane
for 1 hour at room temperature in blocking solution.
Incubate the membrane overnight at 4°C in antibody solution containing a 1:1000 dilution of
anti-PAR rabbit polyclonal antibody. Empirical determination of primary antibody
concentration will be required for optimal results.
Wash the membrane at room temperature for 5 minutes with 3 changes of TBS-0.1%
Tween. Changing the membrane containers often reduces background.
Incubate the membrane at room temperature for 1 hour in antibody solution containing antirabbit
conjugated to horseradish peroxidase.
Empirical determination of secondary antibody concentration will be required for optimal
results.
Wash the membrane for 5 minutes with 4 changes of TBS-0.1% Tween.
Develop peroxidase reaction using chemiluminescence.
- Affar, E.B., et al. 1998. Immunodot blot method for the detection of poly(ADP-ribose) synthesized in vitro and in vivo. Anal Biochem 259:280-3.
- Shah, G.M., et al. 1995. Methods for biochemical study of poly(ADP-ribose) metabolism in vitro and in vivo. Anal Biochem 227:1-13.
- Gagné, J-P., et al. 2008. Proteome-wide identification of poly(ADP-ribose) binding proteins and poly(ADP-ribose)-associated protein complexes. Nucleic Acids Res 36:6959-76.
Citations for PAR/pADPr Antibody
R&D Systems personnel manually curate a database that contains references using R&D Systems products. The data collected includes not only links to publications in PubMed, but also provides information about sample types, species, and experimental conditions.
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Citations: Showing 1 - 8
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Deletion of Topoisomerase 1 in excitatory neurons causes genomic instability and early onset neurodegeneration
Authors: G Fragola, AM Mabb, B Taylor-Bla, JK Niehaus, WD Chronister, H Mao, JM Simon, H Yuan, Z Li, MJ McConnell, MJ Zylka
Nat Commun, 2020-04-23;11(1):1962.
Species: Mouse
Sample Types: Tissue Homogenates
Applications: Western Blot -
The nuclear structural protein NuMA is a negative regulator of 53BP1 in DNA double-strand break repair
Authors: N Salvador M, J Liu, KM Haas, LL Parker, C Chakrabort, SJ Kron, K Hodges, LD Miller, C Langefeld, PJ Robinson, SA Lelièvre, PA Vidi
Nucleic Acids Res., 2019-04-08;0(0):.
Species: Human
Sample Types: Nuclear Extract
Applications: Western Blot -
An uncharacterized FMAG_01619 protein from Fusobacterium mortiferum ATCC 9817 demonstrates that some bacterial macrodomains can also act as poly-ADP-ribosylhydrolases
Authors: AG García-Sau, R Zapata-Pér, JF Hidalgo, J Cabanes, F Gil-Ortiz, Á Sánchez-Fe
Sci Rep, 2019-03-01;9(1):3230.
Species: Human
Sample Types: Recombinant Protein
Applications: Western Blot -
NR4A Nuclear Receptors Target Poly-ADP-Ribosylated DNA-PKcs Protein to Promote DNA Repair
Authors: D Munnur, J Somers, G Skalka, R Weston, R Jukes-Jone, M Bhogadia, C Dominguez, K Cain, I Ahel, M Malewicz
Cell Rep, 2019-02-19;26(8):2028-2036.e6.
Species: Human
Sample Types: Cell Extracts
Applications: Western Blot -
RANKL coordinates multiple osteoclastogenic pathways by regulating expression of ubiquitin ligase RNF146.
Authors: Matsumoto Y, Larose J, Kent O, Lim M, Changoor A, Zhang L, Storozhuk Y, Mao X, Grynpas M, Cong F, Rottapel R
J Clin Invest, 2017-03-13;127(4):1303-1315.
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Cultured networks of excitatory projection neurons and inhibitory interneurons for studying human cortical neurotoxicity.
Authors: Xu J, Fan J, Wang X, Eacker S, Kam T, Chen L, Yin X, Zhu J, Chi Z, Jiang H, Chen R, Dawson T, Dawson V
Sci Transl Med, 2016-04-06;8(333):333ra48.
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The 3'-5' DNA exonuclease TREX1 directly interacts with poly(ADP-ribose) polymerase-1 (PARP1) during the DNA damage response.
Authors: Miyazaki T, Kim Y, Yoon J, Wang H, Suzuki T, Morse H
J Biol Chem, 2014-10-02;289(47):32548-58.
Species: Human
Sample Types: Cell Lysates
Applications: Western Blot -
NuMA promotes homologous recombination repair by regulating the accumulation of the ISWI ATPase SNF2h at DNA breaks.
Authors: Vidi P, Liu J, Salles D, Jayaraman S, Dorfman G, Gray M, Abad P, Moghe P, Irudayaraj J, Wiesmuller L, Lelievre S
Nucleic Acids Res, 2014-04-20;42(10):6365-79.
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