Technical Data
P3112-98
Poly ADP-Ribose (PAR)
Description:
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 PAR is detectable in the first 10 minutes and disappears later on. In keratinocytes MAb 6D639 has been shown to detect UVB-induced apoptosis as early as 4h 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.

Applications:
Suitable for use in Flow Cytometry, Immunocytochemistry, Immunohistochemistry and Western Blot. Other applications have not been tested.

Recommend Dilutions:
Immunocytochemistry: 5-20ug/ml
Immunohistochemistry (Paraffin): 5-20ug/ml. Dilute with 5% non-fat dried milk in PBS.
Western Blot: 2.5ug/ml. Incubate in PBS, 0.05% Tween-20, 5% non-fat dried milk.
Optimal dilutions to be determined by the researcher.

Storage and Stability:
May be stored at 4°C. For long-term storage, aliquot and store at 4°C. Do not freeze. Aliquots are stable for 12 months after receipt. For maximum recovery of product, centrifuge the original vial prior to removing the cap. Further dilutions can be made in assay buffer.
TypeIsotypeCloneGrade
MabIgG36D639Affinity Purified
SizeStorageShippingSourceHost
100ul4°C Do Not FreezeBlue IceMouse
Concentration:
~1mg/ml
Immunogen:
Purified poly(ADP-ribose).
Purity:
Purified by Protein A affinity chromatography
Form
Supplied as a liquid in 50mM HEPES, pH 7.4, 100mM sodium chloride, 1% BSA, 0.02% sodium azide
Specificity:
Recognizes poly(ADP-ribose) synthesized by a broad range of PARPs (poly(ADP-ribose) polymerases) Species Crossreactivity: human, mouse, rat and Drosophila.
Intended for research use only. Not for use in human, therapeutic, or diagnostic applications.
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