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Poly(ADP-ribose) monoclonal antibody (10H)

 
ALX-804-220-R100 100 µl 367.00 USD
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The monoclonal antibody 10H is directed against poly(ADP-ribose) (PAR). PAR is synthesized after activation of the nuclear DNA repair enzyme poly(ADP-ribose)polymerase (PARP). PARP is selectively activated by DNA strand breaks to catalyze the addition of long branched chains of PAR to a variety of nuclear proteins, most notably PARP itself.The amount of PAR formed in living cells with DNA damage is commensurate with the extent of the damage. Under DNA damage conditions, PAR undergoes a rapid turnover, with a half-life in the range of minutes, as PAR is rapidly hydrolyzed and converted to free ADP-ribose by the enzyme poly(ADP-ribose)glycohydrolase (PARG). After massive DNA damage (e.g. γ-irradiation or oxidative stress) PAR is detectable in the first 10 minutes and disappears later on. In keratinocytes MAb 10H has been shown to detect UVB-induced apoptosis as early as 4 hour after irradiation, thus being superior to DNA laddering and the TUNEL assay.Due to the very large number of endonuclease-mediated DNA breaks in apoptosis, PARP becomes strongly activated during the so-called execution phase. In the case of DNA damage-induced apoptosis, this represents a "second round" of PAR synthesis. PAR synthesized during apoptosis appears to be remarkably stable. PAR immunofluorescence appears at least as early during apoptosis as does the specific cleavage of PARP by caspase-3. As shown by several groups, this PAR immunofluorescence correlates well with other markers of apoptosis. MAb to Poly(ADP-ribose) (10H) can be used in flow cytometry.A quantitative non-isotopic immuno-dot-blot method for the assessment of cellular poly(ADP-ribosyl)ation capacity using MAb to Poly(ADP-ribose) (10H) has been described.

Product Specification

Alternative Name:PAR
 
Clone:10H
 
Host:Mouse
 
Isotype:IgG3
 
Immunogen:Purified poly(ADP-ribose).
 
Species reactivity:Human, Mouse, Rat
Drosophila
 
Specificity:Recognizes poly(ADP-ribose) synthesized by a broad range of PARPs (poly(ADP-ribose) polymerases) like human, mouse, rat or Drosophila PARP enzyme.
 
Applications:Flow Cytometry, ICC, IHC (PS), WB
 
Recommended Dilutions/Conditions:Immunocytochemistry (5-20µg/ml)
Immunohistochemistry (paraffin sections; dilution buffer: 5% milk (non fat dried milk) in PBS to a final concentration of 5-20µg/ml)
Western Blot (incubate 2.5µg/ml in PBS, 0.05% Tween20, 5% milk (non fat dried milk))
Suggested dilutions/conditions may not be available for all applications.
Optimal conditions must be determined individually for each application.
 
Purity Detail:Protein A-affinity purified from supernatant.
 
Formulation:Liquid. In 50mM HEPES, pH 7.4, containing 100mM sodium chloride, 1% BSA and 0.02% sodium azide.
 
Handling:Avoid freeze/thaw cycles.
 
Shipping:Shipped on Blue Ice
 
Long Term Storage:-20°C
 
804-220-1
Figure 1: Detection of DNA damage.
804-220-2
Figure 2: Detection of apoptotic cells by immunocytochemistry.
10H Laser
Figure 3: HeLa irradiated cells with a microbeam laser. Picture courtesy of C.Spenlehauer & G. de Murcia (CNRS, Strasbourg)
804-220
Figure 4: Stimulation of PARP activity in permeabilized human PBMC by addition of NAD and activator oligonucleotide, and inhibitory effect of 3-aminobenzamide [21].
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804-220-1 804-220-2 10H Laser 804-220

Product Literature References

HYDAMTIQ, a selective PARP-1 inhibitor, improves bleomycin-induced lung fibrosis by dampening the TGF-β/SMAD signalling pathway: L. Licarini, et al.; J. Cell Mol. Med. (2016), Application(s): Western blot analysis for PARylated protein content, lung tissue homogenates, Abstract; Full Text
17-beta estradiol inhibits oxidative stress-induced accumulation of AIF into nucleolus and PARP1-dependent cell death via estrogen receptor alpha: E. Batnasan, et al.; Toxicol. Lett. 232, 1 (2015), Application(s): Immunocytochemistry using human breast adenocarcinoma cells MCF7, Abstract; Full Text
Adalimumab Reduces Photoreceptor Cell Death in A Mouse Model of Retinal Degeneration: C. Martínez-Fernández de la Cámara, et al.; Sci. Rep. 5, 11764 (2015), Application(s): Immunohistochemistry, Abstract; Full Text
Combinative effects of β-Lapachone and APO866 on pancreatic cancer cell death through reactive oxygen species production and PARP-1 activation: C.S. Breton, et al.; Biochimie 116, 141 (2015), Application(s): Western Blot, Abstract;
Interplay between histone acetylation/deacetylation and poly(ADP-ribosyl)ation in the development of ischemic tolerance in vitro: E. Gerace, et al.; Neuropharmacology 92, 125 (2015), Application(s): Western Blotting, Abstract;
Nuclear-translocated Glyceraldehyde-3-phosphate Dehydrogenase Promotes Poly(ADP-ribose) Polymerase-1 Activation during Oxidative/Nitrosative Stress in Stroke: H. Nakajima, et al.; J. Biol. Chem. 290, 14493 (2015), Abstract; Full Text
PARP is activated in human asthma and its inhibition by olaparib blocks house dust mite-induced disease in mice: M.A. Ghonim, et al.; Clin. Sci. (Lond). 129, 951 (2015), Application(s): Immunoblot analysis, Abstract; Full Text
Two stages of XRCC1 recruitment and two classes of XRCC1 foci formed in response to low level DNA damage induced by visible light, or stress triggered by heat shock: K.J. Solarcyk, et al.; DNA Repair (Amst.) 37, 12 (2015), Application(s): Immunofluorescence, Abstract;
7-Azaindole-1-carboxamides as a new class of PARP-1 inhibitors: R. Cincinelli, et al.; Bioorg. Med. Chem. 22, 1089 (2014), Application(s): Immunocytochemistry using human HeLa cervical carcinoma cells, Abstract;
Erythropoietin Exerts a Neuroprotective Function Against Glutamate Neurotoxicity in Experimental Diabetic Retina: L. Gu, et al.; Invest. Ophthalmol. Vis. Sci. 55, 8208 (2014), Application(s): Immunohistochemistry using rat retina cryosections, Abstract; Full Text
Characterization of stress response in human retinal epithelial cells: V. Giansanti, et al.; J. Cell. Mol. Med. 17, 103 (2013), Application(s): ICC on human adult retinal pigmented epithelial (ARPE-19) cells, Abstract; Full Text
Poly(ADP-Ribose) Polymerase 1 Participates in the Phase Entrainment of Circadian Clocks to Feeding: G. Asher, et al.; Cell 142, 943 (2010), Application(s): WB using mouse liver nuclear extract, Abstract; Full Text
Aldosterone-induced endothelial dysfunction of rat aorta: role of poly(ADP-ribose) activation: A. Tasatargil, et al.; J. Renin Angiotensin Aldosterone Syst. 10, 127 (2009), Abstract;
Substrate-assisted catalysis by PARP10 limits its activity to mono-ADP-ribosylation: H. Kleine, et al.; Mol. Cell 32, 57 (2008), Abstract;
Critical role of inducible nitric oxide synthase in degeneration of retinal capillaries in mice with streptozotocin-induced diabetes: L. Zheng, et al.; Diabetologia 50, 1987 (2007), Abstract;
Flow-cytometric assessment of cellular poly(ADP-ribosyl)ation capacity in peripheral blood lymphocytes: A. Kunzmann, et al.; Immun. Ageing 3, 8 (2006), Application(s): Flow Cytometry, Abstract;
Nuclear poly(ADP-ribose) polymerase-1 rapidly triggers mitochondrial dysfunction: G. Cipriani, et al.; J. Biol. Chem. 280, 17227 (2005), Abstract; Full Text
Activation and Caspase-mediated Inhibition of PARP: A Molecular Switch between Fibroblast Necrosis and Apoptosis in Death Receptor Signaling: M. Los, et al.; Mol. Biol. Cell. 13, 978 (2002), Application(s): Detection of Apoptosis, Abstract; Full Text
Detection of poly(ADP-ribose) by immunocytochemistry: a sensitive new method for the early identification of UVB- and H2O2-induced apoptosis in keratinocytes: H. Chang, et al.; Biol. Chem. 383, 703 (2002), Application(s): Detection of Apoptosis, Abstract;
Poly(ADP-ribose) polymerase cleavage during apoptosis: when and where?: C. Soldani, et al.; Exp. Cell Res. 269, 193 (2001), Application(s): Detection of Apoptosis, Abstract;
Poly(ADP-ribosyl)ation, genomic instability, and longevity: A. Bürkle; Ann. N. Y. Acad. Sci. 908, 126 (2000), Abstract;
Protection against hemorrhagic shock in mice genetically deficient in poly(ADP-ribose)polymerase: L. Liaudet, et al.; PNAS 97, 10203 (2000), Application(s): Immunohistochemistry, Abstract; Full Text
4-Amino-1,8-naphthalimide: a novel inhibitor of poly(ADP-ribose) polymerase and radiation sensitizer: A. Schlicker, et al.; Int. J. Radiat. Biol. 75, 91 (1999), Application(s): Detection of DNA-Damage, Abstract;
Detection of poly(ADP-ribose) synthesis in Drosophila testes upon gamma-irradiation: S. Lankenau, et al.; Chromosoma 108, 44 (1999), Application(s): Detection of DNA-Damage, Abstract;
Overexpression of dominant negative PARP interferes with tumor formation of HeLa cells in nude mice: evidence for increased tumor cell apoptosis in vivo: M.A. Hans, et al.; Oncogene 18, 7010 (1999), Abstract;
Poly(ADP-ribose) immunostaining to detect apoptosis induced by a neurotoxic fragment of prion protein: A. Bürkle, et al.; Histochem. J. 31, 711 (1999), Application(s): Detection of Apoptosis, Abstract;
Quantitative nonisotopic immuno-dot-blot method for the assessment of cellular poly(ADP-ribosyl)ation capacity: R. Pfeiffer, et al.; Anal. Biochem. 275, 118 (1999), Application(s): Immuno-Dot-Blot Detection, Abstract;
Reactive oxygen species participate in mdr1b mRNA and P-glycoprotein overexpression in primary rat hepatocyte cultures: C. Ziemann, et al.; Carcinogenesis 20, 407 (1999), Abstract; Full Text
Selective loss of poly(ADP-ribose) and the 85-kDa fragment of poly(ADP- ribose) polymerase in nucleoli during alkylation-induced apoptosis of HeLa cells: R. Alvarez-Gonzalez, et al.; J. Biol. Chem. 274, 32122 (1999), Application(s): Detection of Apoptosis, Abstract; Full Text
Multiparametric staining to identify apoptotic human cells: C. Negri, et al.; Exp. Cell Res. 234, 174 (1997), Application(s): Detection of Apoptosis, Abstract;
Poly(ADP-ribose) synthesis: a useful parameter for identifying apoptotic cells: M. Donzelli, et al.; Histochem. J. 29, 831 (1997), Application(s): Detection of Apoptosis, Abstract;
trans-dominant inhibition of poly(ADP-ribosyl)ation sensitizes cells against g-irradiation and N-methyl-N'-nitro-N-nitrosoguanidine but does not limit DNA replication of a polyomavirus replicon: J.H. Küpper, et al.; Mol. Cell. Biol. 15, 3154 (1995), Application(s): Detection of DNA-Damage, Abstract; Full Text
Inhibition of poly(ADP-ribosyl)ation by overexpressing the poly(ADP-ribose) polymerase DNA-binding domain in mammalian cells: J.H. Kupper et al.; J. Biol. Chem. 265, 18721 (1990), Abstract;
Rapid assay of poly(ADP-ribose) glycohydrolase: L. Menard & G.G. Poirier; Biochem. Cell Biol. 65, 668 (1987), Abstract;
Monoclonal antibodies to poly(adenosine diphosphate ribose) recognize different structures: H. Kawamitsu, et al.; Biochemistry 23, 3771 (1984), (Original Reference), Abstract;

General Literature References

Poly(ADP-ribose) polymerase and aging: A. Bürkle; Exp. Gerontol. 33, 519 (1998), Abstract;
Detection of poly(ADP-ribose) polymerase and its reaction product poly(ADP-ribose) by immunocytochemistry: J.H. Küpper, et al.; Histochem. J. 28, 391 (1996), Abstract;
Inactivation of the poly(ADP-ribose) polymerase gene affects oxygen radical and nitric oxide toxicity in islet cells: B. Heller, et al.; J. Biol. Chem. 270, 11176 (1995), Abstract; Full Text
Increased poly(ADP-ribosyl)ation in intact cells by cisplatin treatment: A. Bürkle, et al.; Carcinogenesis 14, 559 (1993), Abstract;

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