ADP-ribose-pNP

ADP-ribose-pNP is a colorimetric substrate for assessing activity of poly(ADP-ribose)polymerase (PARP) enzymes. The absorbance of released p-nitrophenol is determined at 405 nm, and the slope of the calibration curve is used to convert the absorbencies to moles of product generated. With ADP-ribose-pNP as the colorimetric substrate, PARP-1 was determined to have the largest Km and Vmax values (151 uM and 1.30 nmolmin-1mg-1 respectively) followed by tankyrase-1 (82 uM and 18 pmolmin-1mg-1 respectively) and VPARP (46 uM and 2 pmolmin-1mg-1 respectively). This colorimetric substrate can be used to determine the kinetic parameters for PARP-1, tankyrase-1, and VPARP, and to screen small-molecule inhibitors of PARP-1, tankyrase-1, and VPARP. ADP-ribose-pNP-based continuous assay has considerable advantages over standard discontinuous PARP assays, enabling the high throughput screening of PARP-1, tankyrase-1, and VPARP activities and their inhibitors.

Protocol

SDS

COA

Catalog	Size	Price	Quantity
11700	1 mg	Price

Calculators

Common stock solution preparation

Table 1. Volume of Water needed to reconstitute specific mass of ADP-ribose-pNP to given concentration. Note that volume is only for preparing stock solution. Refer to sample experimental protocol for appropriate experimental/physiological buffers.

	0.1 mg	0.5 mg	1 mg	5 mg	10 mg
1 mM	138.051 µL	690.255 µL	1.381 mL	6.903 mL	13.805 mL
5 mM	27.61 µL	138.051 µL	276.102 µL	1.381 mL	2.761 mL
10 mM	13.805 µL	69.025 µL	138.051 µL	690.255 µL	1.381 mL

Molarity calculator

Enter any two values (mass, volume, concentration) to calculate the third.

Mass (Calculate)		Molecular weight		Volume (Calculate)		Concentration (Calculate)		Moles
mg	÷	724.37 g/mol	=	mL	x	mM	=	mmol

References

View all 9 references: Citation Explorer

A colorimetric substrate for poly(ADP-ribose) polymerase-1, VPARP, and tankyrase-1

Authors:

Nottbohm AC, Dothager RS, Putt KS, Hoyt MT, Hergenrother PJ.

Journal:

Angew Chem Int Ed Engl (2007): 2066

Poly(ADP-ribose) polymerase as a key player in excitotoxicity and post-ischemic brain damage

Authors:

Meli E, Pangallo M, Baronti R, Chiarugi A, Cozzi A, Pellegrini-Giampietro DE, Moroni F.

Journal:

Toxicol Lett (2003): 153

Pharmacological identification of P2X1, P2X4 and P2X7 nucleotide receptors in the smooth muscles of human umbilical cord and chorionic blood vessels

Authors:

Valdecantos P, Briones R, Moya P, Germain A, Huidobro-Toro JP.

Journal:

Placenta (2003): 17

Kinetic mechanism of nucleotide cofactor binding to Escherichia coli replicative helicase DnaB protein. stopped-flow kinetic studies using fluorescent, ribose-, and base-modified nucleotide analogues

Authors:

Bujalowski W, Jezewska MJ.

Journal:

Biochemistry (2000): 2106

Interactions of nucleotide cofactors with the Escherichia coli replication factor DnaC protein

Authors:

Galletto R, Rajendran S, Bujalowski W.

Journal:

Biochemistry (2000): 12959

Physical properties

Molecular weight	724.37
Solvent	Water

Storage, safety and handling

H-phrase	H303, H313, H333
Hazard symbol	XN
Intended use	Research Use Only (RUO)
R-phrase	R20, R21, R22
Storage	Freeze (< -15 °C); Minimize light exposure
UNSPSC	12171501

Instrument settings

Absorbance microplate reader
Absorbance	405 nm
Recommended plate	Solid white

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Page updated on October 13, 2025