Bovine spleen phosphodiesterase

Manufactured by Merck Group

Sourced in Germany

Bovine spleen phosphodiesterase is an enzyme isolated from bovine spleen tissue. It catalyzes the hydrolysis of phosphodiester bonds in various substrates.

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Lab products found in correlation

Formic acid, by Carl Roth (2 mentions) Calf intestine alkaline phosphatase, by Merck Group (2 mentions) Hplc grade methanol, by Carl Roth (2 mentions) 2 propanol, by Carl Roth (2 mentions) Acetic acid, by Carl Roth (2 mentions) 1 butanol, by Carl Roth (1 mentions) Proteinase k, by Carl Roth (1 mentions) Herring sperm dna, by Merck Group (1 mentions) Micrococcal nuclease from staphylococcus aureus, by Merck Group (1 mentions) Proteinase k, by Qiagen (1 mentions) Agilent 1260 infinity, by Agilent Technologies (1 mentions) Alkaline phosphatase, by Merck Group (1 mentions)

4 protocols using bovine spleen phosphodiesterase

Enzymatic Nucleic Acid Manipulation Protocol

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Calf
intestine alkaline phosphatase, micrococcal
nuclease (from Staphylococcus aureus), bovine spleen
phosphodiesterase, and ribonuclease A from bovine pancreas (RNase)
were purchased from Sigma-Aldrich (Steinheim, Germany). Proteinase
K, HPLC-grade methanol, 2-propanol, 1-butanol, formic acid, and acetic
acid were from Carl Roth GmbH (Karlsruhe, Germany). Herring sperm
DNA and all other reagents and solvents (analytical grade) were from
Sigma-Aldrich.

Monien B.H., Schumacher F., Herrmann K., Glatt H., Turesky R.J, & Chesné C. (2014). Simultaneous Detection of Multiple DNA Adducts in Human Lung Samples by Isotope-Dilution UPLC-MS/MS. Analytical Chemistry, 87(1), 641-648.

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Oligonucleotide Stability Against 5'-Exonuclease

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The stability of oligonucleotides against 5′-exonuclease-mediated degradation was determined using phosphodiesterase II, bovine spleen phosphodiesterase (vial of 10 units, purified, Sigma-Aldrich). The enzyme was dissolved in water according to the manufacturer’s protocol. The assay was performed as described for 3′-exonuclease (vide supra), with the following differences: the oligonucleotide (1.4 nmol, 7 μL of 200 nM stock solution in water) was mixed with acetate buffer (100 mM ammonium acetate, 1 mM EDTA, 1 mM TWEEN 80, pH 6-7) to a volume of 97 μL and cooled down to 0 °C. bovine spleen phosphodiesterase (30 mU, 3 μL of 10 mU/μL stock solution) was then added.

Meng M., Schmidtgall B, & Ducho C. (2018). Enhanced Stability of DNA Oligonucleotides with Partially Zwitterionic Backbone Structures in Biological Media. Molecules, 23(11), 2941.

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Preparation of Isotope-Labeled DNA Adduct

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FFA (CAS: 98-00-0) of 98% purity (Sigma-Aldrich, Steinheim, Germany) was diluted in saline shortly before use. Proteinase K and ribonuclease (RNase) A were purchased from Qiagen (Hilden, Germany). Calf intestine alkaline phosphatase, micrococcal nuclease (from Staphylococcus aureus) and bovine spleen phosphodiesterase were purchased from Sigma-Aldrich. HPLC-grade methanol, 2-propanol, formic acid and acetic acid were from Carl Roth GmbH (Karlsruhe, Germany). The syntheses of the isotope-labelled reference standard [¹⁵N₅,¹³C₁₀]N²-MF-dG has been described previously (22 (link)).

Høie A.H., Monien B.H., Sakhi A.K., Glatt H., Hjertholm H, & Husøy T. (2015). Formation of DNA adducts in wild-type and transgenic mice expressing human sulfotransferases 1A1 and 1A2 after oral exposure to furfuryl alcohol. Mutagenesis, 30(5), 643-649.

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Quantifying DNA Methylation in Cerebellum

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As described before, DNA was extracted from frozen cerebellum tissue via phenol/chloroform/isoamylalcohol extraction followed by enzymatic hydrolysis to obtain respective nucleosides by use of micrococcal nuclease from Staphylococcus aureus, bovine spleen phosphodiesterase and alkaline phosphatase (all Sigma-Aldrich).^{21 (link)} Quantification of methylated (mdC) and hydroxymethylated (hmdC) cytidine in total DNA was conducted on high pressure liquid chromatography (HPLC)–MS/MS (HPLC: Agilent 1260 Infinity, MS/MS: Agilent 6495A).

Friese S., Ranzini G., Tuchtenhagen M., Lossow K., Hertel B., Pohl G., Ebert F., Bornhorst J., Kipp A.P, & Schwerdtle T. (2024). Long-term suboptimal dietary trace element supply does not affect trace element homeostasis in murine cerebellum. Metallomics: Integrated Biometal Science, 16(2), mfae003.

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