Nuclease P1 CAS 54576-84-0

CAS Number

54576-84-0

Grade

Molecular Biology Grade

Molecular Weight

4250

EU Commodity Code

38220090

Shipping Temp

Blue Ice

Storage Temp

-20°C

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

Nuclease P1 (Nuclease 5'-Nucleotidehydrolase, 3'-Phosphohydrolase) from Penicillium citrinum, a zinc dependent glyco-enzyme consisting of 270 amino acid residues, hydrolyzes both 3'-5'-phosphodiester bonds in RNA and heat denatured DNA and 3'-phosphomonoester bonds in mono- and oligonucleotides terminated by 3'-phosphate without base specificity. Nuclease P1 is capable of hydrolyzing single stranded DNA and RNA completely to the level of mononucleoside 5’-monophosphates. The enzyme does not attack double-stranded nucleic acids, especially in the presence of more than 400mM of sodium chloride at pH 6.0.

Activity

≥300U/mg protein using RNA substrate

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°C, pH 5.3. One unit of the enzyme liberates 1umole of orthophosphate from 3’-AMP per minute at 37°C, pH 7.2

Optimum Temperature

~70°C [< 60°C for long incubations]

Stable pH

5.0-8.0

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.

Storage and Stability

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.

Source

Penicillium citrinum

Purity

> 300U/mg protein

Form

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.

Important Note

This product as supplied is intended for research use only, not for use in human, therapeutic or diagnostic applications without the expressed written authorization of United States Biological.

References

General References: 1. Fujimoto, M., et al., Agricultural Biological Chemistry 38: 777-783 (1974). 2. Schomburg's ENZYME HANDBOOK for nuclease P1 from P. citrinum. 3. Shishido, K., Ando, T., Cold Spring Harbor Monograph Series 14: 155-185 (1982). 4. Lehman, R., The Enzymes (Boyer, ed.) 14: 193-201 (1981). 5. Martin, S.A., et al., Biochim. Biophys. Acta 867: 76-80 (1986). 6. “Photochemical Reaction of 7,12-Dimethylbenz[a]anthracene (DMBA) and Formation of DNA Covalent Adducts”, Hongtao Yu,1,* Jian Yan,1,2 Yuguo Jiao,3 and Peter P. Fu2, Int J Environ Res Public Health. (2005) May; 2(1): 114–122. Published online 2005 Apr 30

USBio References

1. Grant, G.P.G., et al., BBRC 371: 451-455 (2008). 2. Yang, Z., et al., Chem. Res. Toxicol. 18: 1339–1346 (2005). 3. McLean, L., et al., Int. J. Cancer 122: 1665-1674 (2007). 4. González-Espinosa, D., et al., Int. Wound J. 4: 241-250 (2007). 5. Dong, H., Drug Metabolism & Disposition 34: 1122-1127 (2006). 6. Jaruga, P. & Dizdaroglu, M., BBRC 397: 48-52 (2010). 7. Kirkali, G. et al., DNA Repair 8: 274-278 (2009). 8. Winitsky, S.O., et al., PLoS Biol. 3: e87 (2005). 9. Jaruga, P., et al., BBRC 386: 656-660 (2009). 10. Rodriguez, H., et al., Biochemistry 46: 2488-2496 (2007). 11. Kim, S.Y., et al., Chem. Res. Toxicol. 19: 852-858 (2009). 12. Nelson, B.C., et al., (2011) Nanotoxicology, doi:10.3109/17435390.2011.626537. 13. Barzideh, J. et al., (2012) Andrologia DOI: 10.1111/and.12033. 14. Buhimschi, A.D. and Gasparro, F. P., (2013) Photochemistry and Photobiology, doi: 10.1111/php.12171. 15. Trotman JB and Schoenberg DR. RNA Cap Methyltransferase Activity Assay. 2018. Bio Protoc. 8(6):2767. doi: 10.21769/BioProtoc.2767. 16. Pang D.,et al. 2014. Significant disparity in base and sugar damage in DNA resulting from neutron and electron irradiation. J Radiat Res. 55(6):1081-1088. 17. Dong X., et al. 2018. Effect of luteolin on the methylation status of the OPCML gene and cell growth in breast cancer cells. Experimental and Therapeutic Medicine. https://doi.org/10.3892/etm.2018.6526

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N7000 Nuclease P1 (Nuclease 5'-Nucleotidehydrolase, 3'-Phosphohydrolase, NP1) CAS: 54576-84-0

CAS Number

Grade

Molecular Weight

EU Commodity Code

Shipping Temp

Storage Temp

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

References

USBio References

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