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Shanghai Qifa Biotechnology Co., Ltd
Thermal instability resistant high salt nuclease (HL-SAN)
NegotiableUpdate on 05/06
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Overview
Heat Labile Salt Active Nuclease (HL-SAN) is a non-specific endonuclease with excellent activity at high salt concentrations; And the enzyme is active in various buffer solutions and is easily deactivated by reducing agent treatment. These characteristics make HL-SAN more suitable for applications that require the removal of DNA and RNA.
Product Details
Product Introduction
nothingWhether it is laboratory scale sample preparation or production scale process, removing nucleic acid can improve the process flow, such as protein purification, virus vector preparation, sample processing in mNGS, etc. The selection of nucleases is particularly important. Heat Labile Salt Active Nuclease (HL-SAN) is a non-specific endonuclease with excellent activity at high salt concentrations; And the enzyme is active in various buffer solutions and is easily deactivated by reducing agent treatment. These characteristics make HL-SAN more suitable for applications that require the removal of DNA and RNA.
Nucleic acid contamination, especially host genome DNA contamination, is a key issue that needs to be addressed in almost all production processes and biological products. On the one hand, the presence of host DNA affects the purification and downstream quality analysis of the target product. On the other hand, residual host DNA can cause severe immune reactions in the body and is one of the key quality parameters of biological products. FDA and NMPA have strict quality control requirements for Host Cell DNA (HCD). In the host genome DNA, histones and DNA form nucleosomes, which fold tightly to form complex chromatin structures. There are strong ionic and hydrophobic interactions between histones and DNA, coupled with their unique structure, making it difficult to accurately detect and eliminate host DNA. Previous literature has shown that histone and DNA dissociation is relatively low at high salt concentrations, which is beneficial for the detection and removal of host DNA. But the vast majority of nucleases available on the market have weak activity or even inactivation at high salt concentrations. And HL-SAN, which is resistant to high salt, has become an ideal choice for such applications.
Product Features
Under high salt conditions, DNA clearance efficiency is higher and there is less residual host DNA;
The pI is 9.6, making it easy to separate from the vast majority of proteins;
When reducing agents are present, enzymes are prone to deactivation, reducing their impact on subsequent processes.
Scope of application
Application of pathogenic microorganism detection, such as metagenomic sequencing (mNGS) to remove host DNA from samples;
Protein purification, especially DNA binding proteins;
Preparation of viral vectors, such as adeno-associated virus (AAV), adenovirus (AdV), etc;
Other applications that require the removal of host DNA, etc.
Precautions
For your safety and health, please wear lab coats and disposable gloves when operating!
This product is mainly used in the field of scientific research and should not be used for clinical diagnosis or other purposes.
quality control
Source: Recombinant production in Pichia pastoris.
Activity: HL-SAN exhibits high activity within the temperature range of 10-50 ° C. The optimal NaCl concentration for the activity is 0.5 M, with a working range of 0.25-1 M. The activity requires Mg2+(>1 mM). The working pH range is 7.5-9.5, with an excellent pH value of 9.0.
Specific activity: ≥ 175000 Units/mg.
Unit definition: One unit is defined as the increase in absorbance of 1 A at 260 nm within 30 minutes at 37 ° C, using 50 μ g/ml calf thymus DNA (D-1501, Sigma) in a buffer composed of 25 mM Tris - HCl, pH 8.5 (25 ° C), 5 mM MgCl, and 2500 mM NaCl.
