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Poly(ADP-ribose) monoclonal antibody (10H)
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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.{{927685-43-6} medchemexpress|{927685-43-6} Technical Information|{927685-43-6} In Vivo|{927685-43-6} supplier} As shown by several groups, this PAR immunofluorescence correlates well with other markers of apoptosis.{{209541-26-4} site|{209541-26-4} Protocol|{209541-26-4} Data Sheet|{209541-26-4} manufacturer} MAb to Poly(ADP-ribose) (10H) can be used in flow cytometry.PMID:25905299 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. Figure 2: Detection of apoptotic cells by immunocytochemistry. Figure 1: Detection of DNA damage. Figure 3: HeLa irradiated cells with a microbeam laser. Picture courtesy of C.Spenlehauer & G. de Murcia (CNRS, Strasbourg) Figure 4: Stimulation of PARP activity in permeabilized human PBMC by addition of NAD and activator oligonucleotide, and inhibitory effect of 3-aminobenzamide [21]. Figure 2: Detection of apoptotic cells by immunocytochemistry. Figure 1: Detection of DNA damage. Figure 3: HeLa irradiated cells with a microbeam laser. Picture courtesy of C.Spenlehauer & G. de Murcia (CNRS, Strasbourg) 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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| Alternative Name PAR | Application Flow Cytometry, ICC, IHC (PS), WB | Clone 10H | Formulation Liquid. In 50mM HEPES, pH 7.4, containing 100mM sodium chloride, 1% BSA and 0.02% sodium azide. | Host Mouse | Immunogen Purified poly(ADP-ribose). | Isotype IgG3 | Recommendation 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. | Species Reactivity Drosophila, Human, Mouse, Rat | Specificity Recognizes poly(ADP-ribose) synthesized by a broad range of PARPs (poly(ADP-ribose) polymerases) like human, mouse, rat or Drosophila PARP enzyme. | Technical Info / Product Notes Cited samples:For an overview on cited samples please click here. | Unit of Measure (UM) µl
