| Size | Price | Stock | Qty |
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| 1mg |
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| Other Sizes |
| Targets |
The compound targets the enzymatic activity of Poly(ADP-ribose) glycohydrolase (PARG) and ADP-ribosyl hydrolase 3 (ARH3). These enzymes are responsible for cleaving the glycosidic bond between the ribose and ADP-ribose moieties of poly(ADP-ribose) (PAR) generated by PARP activity. The substrate is also useful for general poly(ADP-ribose)polymerase (PARP) enzyme studies. By interacting with the active site of these hydrolases, pNP-ADPr serves as a tool to probe the catalytic mechanisms of PAR hydrolysis without acting as a traditional ligand or drug target.
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| ln Vitro |
In the initial continuous Poly(ADP-ribose) glycohydrolase (PARG) and ADP-ribosyl hydrolase 3 (ARH3) activity tests, pNP-ADPr is the colorimetric substrate employed[1].
In vitro, pNP-ADPr disodium acts as a chromogenic substrate for the first continuous assays measuring PARG and ARH3 activity. Upon enzymatic cleavage of the ADP-ribose-pNP bond, the p-nitrophenol group is released, generating a yellow color that can be quantified spectrophotometrically at 405 nm. This allows for real-time, kinetic measurement of enzyme activity, eliminating the need for radioactive substrates. It is also used in the study of PARP enzymes. |
| ln Vivo |
In vivo activity data for pNP-ADPr disodium have not been reported or are not applicable for this compound. As a substrate-based research tool designed for in vitro biochemical assays, pNP-ADPr disodium is not intended for or used in animal model studies. Therefore, no information regarding its in vivo efficacy, pharmacokinetics, or pharmacodynamics in living organisms is currently available in the scientific literature or product documentation.
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| Enzyme Assay |
A typical continuous colorimetric assay protocol: Prepare reaction buffer (50 mM Tris-HCl, pH 7.5, 5 mM MgCl2, 1 mM DTT) in a 96-well plate. Add purified PARG or ARH3 enzyme (0.1-1 microg) to wells containing pNP-ADPr disodium (100-500 microM final concentration). Immediately monitor absorbance at 405 nm every 30 seconds for 30 minutes at 37degC using a microplate reader. The increase in absorbance at 405 nm, corresponding to p-nitrophenol release, is proportional to enzyme activity. A control well without enzyme is used to subtract background. No pre-incubation period is required. Calibration curves can be generated using known concentrations of p-nitrophenol. Enzyme kinetic parameters (Vmax, Km, kcat) are calculated by fitting initial rate data (change in absorbance per minute) to the Michaelis-Menten equation.
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| Cell Assay |
While not a traditional cell-based assay, the compound is used in cell lysate assays to quantify PARG/ARH3 activity. Cells are harvested and lysed in ice-cold lysis buffer (50 mM Tris-HCl, pH 7.5, 150 mM NaCl, 1% NP-40, protease inhibitors). Lysates are cleared by centrifugation at 12,000 × g for 10 min. Protein concentration is normalized using a BCA assay. 20-50 microg of lysate protein is added to each well of a 96-well plate. pNP-ADPr disodium is added to a final concentration of 200 microM. The plate is incubated at 37degC for 30-60 min. Absorbance at 405 nm is measured kinetically or as an endpoint. The measured activity is proportional to the amount of PARG/ARH3 in the lysate. The protocol allows for comparisons between treatment groups (e.g., drug-treated vs. control cells).
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| Animal Protocol |
The compound is not intended for in vivo animal studies, and no standard animal protocol exists for this reagent. If used in an ex vivo context, it can be applied to tissue homogenates following the same lysate-based protocol: homogenize the tissue of interest (e.g., 50-100 mg) in lysis buffer, centrifuge to clear, measure protein concentration, and incubate normalized lysates with pNP-ADPr disodium substrate (200-500 microM) at 37degC for 30-60 min. Measure the absorbance at 405 nm. This approach would measure the total PARG/ARH3 activity extracted from the tissue sample.
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| ADME/Pharmacokinetics |
Pharmacokinetic parameters have not been determined for this compound, as it is an in vitro biochemical reagent not intended for systemic administration. The disodium salt form confers high aqueous solubility (e.g., 250 mg/mL in water), which is an important physicochemical property for its use in buffer systems. However, no data on absorption, distribution, metabolism, elimination, or half-life are available. Researchers are advised to handle this compound as a standard laboratory chemical and not as a potential drug candidate.
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| Toxicity/Toxicokinetics |
No detailed toxicity data are available for pNP-ADPr disodium as it is not intended for in vivo use. Standard laboratory safety precautions should be followed when handling the powder or solutions. Avoid inhalation, ingestion, and skin contact. Use appropriate personal protective equipment (PPE) including gloves, lab coat, and eye protection. The compound should be handled in a well-ventilated area, such as a chemical fume hood. In the event of a spill, follow institutional hazardous material protocols. If skin or eye contact occurs, rinse thoroughly with water for 15 minutes and seek medical advice if irritation persists.
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| References | |
| Additional Infomation |
This product is strictly a research-grade biochemical tool and is not approved for clinical, diagnostic, or therapeutic use in any jurisdiction. It is a colorimetric substrate used for the first continuous PARG/ARH3 activity assays. The presence of the p-nitrophenyl group enables easy spectrophotometric detection. The disodium salt is the standard form to ensure solubility in aqueous buffers. The compound is stable for 3 years when stored as a powder at -20degC and for 2 years at 4degC. In solution, it is stable for 6 months at -80degC. Store in a sealed, protected environment away from moisture and light. This product is for research use only and not for human use.
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| Molecular Formula |
C21H24N6NA2O16P2
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|---|---|
| Molecular Weight |
724.37
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| Related CAS # |
pNP-ADPr;939028-75-8
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| Appearance |
White to off-white solid powder
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| HS Tariff Code |
2934.99.9001
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| Storage |
Powder -20°C 3 years 4°C 2 years In solvent -80°C 6 months -20°C 1 month Note: Please store this product in a sealed and protected environment (e.g. under nitrogen), avoid exposure to moisture and light. |
| Shipping Condition |
Room temperature (This product is stable at ambient temperature for a few days during ordinary shipping and time spent in Customs)
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| Solubility (In Vitro) |
H2O :~250 mg/mL (~345.13 mM)
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| Solubility (In Vivo) |
Note: Listed below are some common formulations that may be used to formulate products with low water solubility (e.g. < 1 mg/mL), you may test these formulations using a minute amount of products to avoid loss of samples.
Injection Formulations
Injection Formulation 1: DMSO : Tween 80： Saline = 10 : 5 : 85 (i.e. 100 μL DMSO stock solution → 50 μL Tween 80 → 850 μL Saline)(e.g. IP/IV/IM/SC) *Preparation of saline: Dissolve 0.9 g of sodium chloride in 100 mL ddH ₂ O to obtain a clear solution. Injection Formulation 2: DMSO : PEG300 ：Tween 80 : Saline = 10 : 40 : 5 : 45 (i.e. 100 μL DMSO → 400 μLPEG300 → 50 μL Tween 80 → 450 μL Saline) Injection Formulation 3: DMSO : Corn oil = 10 : 90 (i.e. 100 μL DMSO → 900 μL Corn oil) Example: Take the Injection Formulation 3 (DMSO : Corn oil = 10 : 90) as an example, if 1 mL of 2.5 mg/mL working solution is to be prepared, you can take 100 μL 25 mg/mL DMSO stock solution and add to 900 μL corn oil, mix well to obtain a clear or suspension solution (2.5 mg/mL, ready for use in animals). View More
Injection Formulation 4: DMSO : 20% SBE-β-CD in saline = 10 : 90 [i.e. 100 μL DMSO → 900 μL (20% SBE-β-CD in saline)] Oral Formulations
Oral Formulation 1: Suspend in 0.5% CMC Na (carboxymethylcellulose sodium) Oral Formulation 2: Suspend in 0.5% Carboxymethyl cellulose Example: Take the Oral Formulation 1 (Suspend in 0.5% CMC Na) as an example, if 100 mL of 2.5 mg/mL working solution is to be prepared, you can first prepare 0.5% CMC Na solution by measuring 0.5 g CMC Na and dissolve it in 100 mL ddH2O to obtain a clear solution; then add 250 mg of the product to 100 mL 0.5% CMC Na solution, to make the suspension solution (2.5 mg/mL, ready for use in animals). View More
Oral Formulation 3: Dissolved in PEG400 (Please use freshly prepared in vivo formulations for optimal results.) |
| Preparing Stock Solutions | 1 mg | 5 mg | 10 mg | |
| 1 mM | 1.3805 mL | 6.9025 mL | 13.8051 mL | |
| 5 mM | 0.2761 mL | 1.3805 mL | 2.7610 mL | |
| 10 mM | 0.1381 mL | 0.6903 mL | 1.3805 mL |
*Note: Please select an appropriate solvent for the preparation of stock solution based on your experiment needs. For most products, DMSO can be used for preparing stock solutions (e.g. 5 mM, 10 mM, or 20 mM concentration); some products with high aqueous solubility may be dissolved in water directly. Solubility information is available at the above Solubility Data section. Once the stock solution is prepared, aliquot it to routine usage volumes and store at -20°C or -80°C. Avoid repeated freeze and thaw cycles.
Calculation results
Working concentration： mg/mL；
Method for preparing DMSO stock solution： mg drug pre-dissolved in μL DMSO (stock solution concentration mg/mL). Please contact us first if the concentration exceeds the DMSO solubility of the batch of drug.
Method for preparing in vivo formulation:：Take μL DMSO stock solution, next add μL PEG300, mix and clarify, next addμL Tween 80, mix and clarify, next add μL ddH2O，mix and clarify.
(1) Please be sure that the solution is clear before the addition of next solvent. Dissolution methods like vortex, ultrasound or warming and heat may be used to aid dissolving.
(2) Be sure to add the solvent(s) in order.
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