Phosphodiesterase 1 Enzyme | PDE1 enzyme
Phosphodiesterase 1
Purity
Molecular Biology Grade
Synonyms
Phosphodiesterase 1; N/A; PDE1 enzyme
Host
Crotalus adamanteus venom (Eastern Diamondback rattlesnake)
Purity/Purification
Molecular Biology Grade
Form/Format
Supplied as a lyophilized powder
Activity
72.8 u/mg dry weight, 110 u/vial
Absorbance (A280)
1.68
Nucleotidase
0.75%
Optimum pH
9.8-10.4
Inhibitors
Reducing agents such as glutathione, cysteine and ascorbic acid. It is completely inhibited by 5mM EDTA while ATP, ADP and AMP are partial inhibitors.
Activators
The enzyme has an absolute requirement for Mg2+ indicates an optimum concentration of 15mM.
Unit Definition
1 unit hydrolyzes 1 umole of p-nitrophenyl thymidine-5-phosphate per minute at 25ºC, pH 8.9.
Protocol
Assay:
Method:
The assay is essentially that of Razell and Khorana (1959) where the reaction velocity is determined by an increase in absorbance at 400nm resulting from the hydrolysis of p-nitrophenyl thymidine -5'-phosphate. One unit hydrolyzes one micromole of p-nitrophenyl thymidine-5'-phosphate per minute at pH 8.9 and 25°C under the specified conditions.
Reagents:
-0.11M Tris-HCl buffer, pH 8.9, with 0.11M sodium chloride and 15 mM MgCl2 (Tris*Salts buffer)
- 5 mM p-nitrophenyl thymidine-5'-phosphate.
Note: The purity of commercial preparations varies somewhat and should be considered in preparing this reagent.
Enzyme:
Dissolve at 1 mg/ml in Tris*Salts buffer to obtain a rate of 0.02-0.04 ΔA/minute.
Protocol:
1. Set spectrophotometer at 400nm and 25ºC.
2. Pipette into microcuvettes as follows:
Tris*Salts buffer 0.9 ml; 5 mM p-nitrophenyl thymidine-5'-phosphate 0.1 ml.
3. Incubate cuvettes in spectrophotometer for 3-5 minutes to reach temperature equillibrium and establish blank rate, if any. Add 10ul of diluted enzyme and record increase in A400 for 3-5 minutes. The reaction remains linear until A400 reaches about 1.2. Calculate ΔA400/minute from initial linear portion of absorbance curve.
Calculation:
Units/mg = ΔA400/min/16* x mg enzyme/ml reaction mixture
*16 is the extinction coefficient of p-nitrophenol determined in the laboratory under these conditions.
Method:
The assay is essentially that of Razell and Khorana (1959) where the reaction velocity is determined by an increase in absorbance at 400nm resulting from the hydrolysis of p-nitrophenyl thymidine -5'-phosphate. One unit hydrolyzes one micromole of p-nitrophenyl thymidine-5'-phosphate per minute at pH 8.9 and 25°C under the specified conditions.
Reagents:
-0.11M Tris-HCl buffer, pH 8.9, with 0.11M sodium chloride and 15 mM MgCl2 (Tris*Salts buffer)
- 5 mM p-nitrophenyl thymidine-5'-phosphate.
Note: The purity of commercial preparations varies somewhat and should be considered in preparing this reagent.
Enzyme:
Dissolve at 1 mg/ml in Tris*Salts buffer to obtain a rate of 0.02-0.04 ΔA/minute.
Protocol:
1. Set spectrophotometer at 400nm and 25ºC.
2. Pipette into microcuvettes as follows:
Tris*Salts buffer 0.9 ml; 5 mM p-nitrophenyl thymidine-5'-phosphate 0.1 ml.
3. Incubate cuvettes in spectrophotometer for 3-5 minutes to reach temperature equillibrium and establish blank rate, if any. Add 10ul of diluted enzyme and record increase in A400 for 3-5 minutes. The reaction remains linear until A400 reaches about 1.2. Calculate ΔA400/minute from initial linear portion of absorbance curve.
Calculation:
Units/mg = ΔA400/min/16* x mg enzyme/ml reaction mixture
*16 is the extinction coefficient of p-nitrophenol determined in the laboratory under these conditions.
Preparation and Storage
Lyophilized and reconstituted products are stable for 6 months after receipt at -20ºC. Reconstitute with sterile dH2O. Aliquot to avoid repeated freezing and thawing. Store at -20ºC. 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.
Related Product Information for PDE1 enzyme
Venom exonuclease (Phosphodiesterase I) successively hydrolyzes 5'-mononucleotides from 3'-hydroxy-terminated ribo-and deoxyribo-oligonucleotides. The enzyme has been widely utilized as a tool for structural and sequence studies of nucleic acids. The enzyme has been purified with endonuclease activity being eliminated as well as 5'-nucleotidase and nonspecific monophosphatase. It is nonspecific with respect to base or sugar moieties of nucleotides. A variety of synthetic substrates are hydrolyzed. The exonuclease will not recognize nucleoside units in the syn conformation. ADP-ribosylated proteins are cleaved at the pyrophosphate linkages by venom phosphodiesterase to yield phosphoribosyl-AMP.
Product Categories/Family for PDE1 enzyme
NCBI and Uniprot Product Information
NCBI GI #
NCBI GeneID
NCBI Accession #
NCBI GenBank Nucleotide #
Molecular Weight
115kD
NCBI Official Full Name
phosphodiesterase 1
NCBI Official Symbol
PDE1
NCBI Protein Information
phosphodiesterase 1
Similar Products
Product Notes
The PDE1 (Catalog #AAA14846) is an Enzyme produced from Crotalus adamanteus venom (Eastern Diamondback rattlesnake) and is intended for research purposes only. The product is available for immediate purchase. It is sometimes possible for the material contained within the vial of "Phosphodiesterase 1, Enzyme" to become dispersed throughout the inside of the vial, particularly around the seal of said vial, during shipment and storage. We always suggest centrifuging these vials to consolidate all of the liquid away from the lid and to the bottom of the vial prior to opening. Please be advised that certain products may require dry ice for shipping and that, if this is the case, an additional dry ice fee may also be required.Precautions
All products in the AAA Biotech catalog are strictly for research-use only, and are absolutely not suitable for use in any sort of medical, therapeutic, prophylactic, in-vivo, or diagnostic capacity. By purchasing a product from AAA Biotech, you are explicitly certifying that said products will be properly tested and used in line with industry standard. AAA Biotech and its authorized distribution partners reserve the right to refuse to fulfill any order if we have any indication that a purchaser may be intending to use a product outside of our accepted criteria.Disclaimer
Though we do strive to guarantee the information represented in this datasheet, AAA Biotech cannot be held responsible for any oversights or imprecisions. AAA Biotech reserves the right to adjust any aspect of this datasheet at any time and without notice. It is the responsibility of the customer to inform AAA Biotech of any product performance issues observed or experienced within 30 days of receipt of said product. To see additional details on this or any of our other policies, please see our Terms & Conditions page.Item has been added to Shopping Cart
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