Thermo Scientific™ Pierce™ Universal-Nuklease für die Zelllyse

Beschreibung

Thermo Scientific Pierce Universal Nuclease for Cell Lysis is an endonuclease that is ideal for a wide variety of applications where complete digestion of nucleic acids is needed when preparing cell lysates.

Features of Universal Nuclease for Cell Lysis include:

• Broad spectrum—degrades all forms of DNA and RNA
• Highest-quality enzyme—nuclease is ≥99% pure, as tested by SDS-PAGE
• Robust activity—100-fold greater specific activity than DNase I
• Versatile—can be used with a wide variety of cell lysis reagents

Pierce Universal Nuclease for Cell Lysis is a genetically engineered endonuclease from Serratia marcescens. The enzyme is produced and purified from E. coli and consists of two identical 30-kDa subunits with two critical disulfide bonds. This indiscriminate endonuclease degrades single-stranded, double-stranded, linear, and circular DNA and RNA and is effective over a wide range of temperatures and pH.

Pierce Universal Nuclease for Cell Lysis has high specific activity (100-fold greater than DNase I) and increased thermal stability compared to other nucleases. It is ≥99% pure enzyme, free of any measurable protease activity, and supplied at 250U/μL. It is identical in performance to Benzonase™ Nuclease (EMD Merck).

Applications

• Use with B-PER, Y-PER, or other commercial or homebrew cell lysis reagents and/or mechanical disruption to reduce viscosity in protein extracts
• Use to remove DNA and RNA from recombinant protein preparations prior to downstream processing

Pierce Universal Nuclease for Cell Lysis is commonly used to reduce the viscosity of bacterial and mammalian protein extracts for downstream application by removing the nucleic acids from protein preparations. The enzyme completely digests nucleic acids to oligonucleotides that are less than five bases long. It helps to improve the separation of the lysate pellet from the supernatant, enhances filtration of the treated lysate, improves chromatography processing time, and increases the overall protein yield. It has also been shown to improve the compatibility of protein extracts for 2D gel electrophoresis.

One unit corresponds to the amount of enzyme required to produce a change of 1.0 in the absorbance at 260 nm of sonicated herring DNA over 30 minutes at 37°C, as determined using standard nuclease from the Merck™ Serratia marcescens volumetric activity assay. Add 25 U, or 0.1 µL, of Pierce Universal Nuclease for Cell Lysis to 1 mL of cell lysate and incubate at room temperature for 10–15 min.