PARP catalyzes the NAD⁺-dependent addition of poly(ADP-ribose) (PAR) onto itself and adjacent nuclear proteins. This enzyme is a therapeutic target for BRCA1 and BRCA2 associated breast cancers. To address the need to monitor PARP activity among different individuals and within cells, Trevigen's improved and validated HT PARP in vivo Pharmacodynamic Assay II measures net PAR levels in tissue or cellular extracts and has been used to document differences in PAR levels among tumor lysates, organs, and xenografts. The HT PARP in vivo Pharmacodynamic Assay II employs a 96 well plate, pre-coated with Trevigen's monoclonal PAR antibody as the capture agent, and anti-PAR polyclonal rabbit antibody as the detecting agent.

FEATURES:

• Pre-coated 96 well capture antibody plates
• High signal to noise ratio
• Detection sensitivity of 2 pg/ml of PAR
• Broad linear dynamic range to 1,000 pg/ml
• Reduced inter-assay variability
• Validated assay that measures drug action on PARP in both in vivo and in vitro settings
• 96 test size

APPLICATIONS:

• Quantitation of PAR in peripheral blood mononuclear cells, tissue culture cells, and tumor lysates from different tissues, organs and xenografts.
• Monitoring the efficacy of PARP inhibitors on PAR formation in vivo.
• Verifying observations of enhanced cancer cell cytotoxicity arising from PARP inhibitor/anticancer drug combination therapy.
• Facilitating development of PARP and PARG targeted therapeutics.

KIT COMPONENTS:

• PAR Standard and Sample Buffer 
• PAR Polyclonal Detecting Ab and Diluent
• Goat anti-Rabbit IgG-HRP 
• Detection Reagents
• Cell Lysis Reagent and 20% SDS 
• DNase I and Mg Cation
• Jurkat Cell Lysate Standard Controls (Low, Medium, and High)
• Pre-coated 96-stripwell plate and sealers 

REFERENCES:

1. Virag, L., Szabo, C. 2002. The therapeutic potential of Poly(ADP-Ribose) Polymerase inhibitors. Pharmacological Reviews 54:375-429.

2. Curtin NJ. 2005. PARP inhibitors for cancer therapy. Expert Rev Mol Med 7:1-20.

3. Kinders JK, Hollingshead M, Khin S, Rubinstein L, Tomaszewski JE, Doroshow JH, Parchment RE, and the National Cancer Institute Phase 0 Clinical Trials Team. 2008. Preclinical Modeling of a Phase 0 Clinical Trial: Qualification of a Pharmacodynamic Assay of Poly (ADP-Ribose) Polymerase in Tumor Biopsies of Mouse Xenografts. Clin Cancer Res 14:6877-85.

4. Bryant HE, Schultz N, Thomas HD, Parker KM, Flower D, Lopez E, Kyle S, Meuth M, Curtin NJ, Helleday T. 2005. Specific killing of BRCA2-deficient tumours with inhibitors of poly(ADP-ribose) polymerase. Nature 434:913-7.

5. Farmer H, McCabe N, Lord CJ, Tutt AN, Johnson DA, Richardson TB, Santarosa M, Dillon KJ, Hickson I, Knights C, Martin NM, Jackson SP, Smith GC, Ashworth A. 2005. Targeting the DNA repair defect in BRCA mutant cells as a therapeutic strategy. Nature 434:917-21.

6. Fong PC, Boss DS, Yap TA, Tutt A, Wu P, Mergui-Roelvink M, Mortimer P, Swaisland H, Lau A, O'Connor MJ, Ashworth A, Carmichael J, Kaye SB, Schellens JH, de Bono JS. 2009. Inhibition of poly(ADP-ribose) polymerase in tumors from BRCA mutation carriers. N Engl J Med 361:123-34.