Nuclease P1 Enzyme

Nuclease P1 (Nuclease 5'-Nucleotidehydrolase, 3'-Phosphohydrolase, NP1)

Cat. Num. AAA14839

• Purity: Molecular Biology Grade
• Synonyms: Nuclease P1; N/A; Nuclease P1 (Nuclease 5'-Nucleotidehydrolase, 3'-Phosphohydrolase, NP1); Nuclease P1 enzyme

• Host: Penicillium citrinum
• Specificity: 410u/mg protein
• Purity/Purification: Molecular Biology Grade
• Form/Format: Supplied as a powder lyophilized from a solution containing 30mM sodium acetate buffer, pH 5.3 with 5mM ZnCl2 and 50mM sodium chloride. Enzyme suspension obtained after reconstitution.
• Concentration: Protein Concentration (before lyophilization): 1.50 mg/ml (varies by lot)

• Specific Activity: 410 u/mg protein
• Stable pH: 5.0-8.0
• Optimum pH: 5.3 for RNA and heat-denatured DNA; 7.2 for 3' -AMP
• Optimum Temperature: ~70°C [<60°C for long incubations]
• Reconstitution: For maximum recovery of product, briefly centrifuge the original vial prior to opening. This will dislodge any enzyme that has adhered to the side or in the cap due to shipping. Reconstitute with sterile ddH2O. Allow the nuclease to rest on ice for ~10-15 minutes. Vortex gently to ensure there is uniform suspension (Nuclease tends to become layered towards the bottom of the vial). Just before addition to the reaction mixture, gently vortex the suspension.
• Unit Definition: One unit of nuclease activity is defined as the amount of enzyme that produces 1umole of acid soluble nucleotides from RNA based on E = 10,600 for RNA hydrolysates per minute at 37 degree C, pH 5.3. One unit of the enzyme liberates 1umole of orthophosphate from 3'-AMP per minute at 37 degree C, pH 7.2
• Substrate specificity: The enzyme hydolyzes both 3'-5' phosphodiester bonds in RNA and heat-denatured DNA, and 3'-phosphomonoester bonds in mono and oligonucleotides terminated with a 3'-phosphate group without base specificity. The enzyme does not attack double-stranded nucleic acids, especially in the presence of more than 400mM of sodium chloride at pH 6.0.
• Preparation and Storage: Lyophilized powder may be stored at -20°C. Stable for 12 months. Reconstituted product should be aliquoted and stored at -20°C. Reconstituted product is stable for 2-3 weeks at 4°C and 3-6 months at -20°C. For maximum recovery of product, centrifuge the original vial after thawing and prior to removing the cap. Further dilutions can be made in assay buffer.

Application Data

(EC=3.1.30.1; Endonuclease P1; Nuclease 5'-Nucleotidehydrolase; 3'-Phosphohydrolase; NP1)

Application Data

(Gen. Ref. 6:Autoradiogram of 32P-postlabeled nuclease P1-treated DNA from photo-irradiation in the presence of (A) DMBA, (B) blank, (C) 7-HOCH2-12-MBA, (D) 12-HOCH2-7-MBA, (E) 7-CHO-12-MBA, and (F) 12-CHO-7-MBA in THF and calf thymus DNA by UVA light at a total dose of 14 J/cm2.)

Application Data

(Nuclease P1 (Nuclease 5'-Nucleotidehydrolase, 3'-Phosphohydrolase, NP1))

Product Categories/Family for Nuclease P1 enzyme

Molecular Biology; MB-Enzymes

NCBI and Uniprot Product Information

• NCBI GI #: 236381
• Molecular Weight: 42-50kD
• NCBI Official Full Name: nuclease P1 {EC 3.1.30.1}